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1.
Nitta, Mai.
Approaches to understanding the mechanism of mammalian folliculogenesis : Analysis of gap junctions in ovarian follicles, and the establishment of an in vitro system for obtaining follicles from mouse embryonic stem cells : 哺乳類卵胞形成機構の解析：卵胞発育におけるギャップ結合の解析とマウス胚性幹細胞からの卵胞誘導系の構築; ホニュウルイ ランホウ ケイセイ キコウ ノ カイセキ ： ランホウ ハツイク ニ オケル ギャップ ケツゴウ ノ カイセキ ト マウス ハイセイカン サイボウ カラノ ランホウ ユウドウケイ ノ コウチク.
Degree: Nara Institute of Science and Technology / 奈良先端科学技術大学院大学
URL: http://hdl.handle.net/10061/5726 
Subjects/Keywords: ES cells
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APA (6th Edition):
Nitta, M. (n.d.). Approaches to understanding the mechanism of mammalian folliculogenesis : Analysis of gap junctions in ovarian follicles, and the establishment of an in vitro system for obtaining follicles from mouse embryonic stem cells : 哺乳類卵胞形成機構の解析：卵胞発育におけるギャップ結合の解析とマウス胚性幹細胞からの卵胞誘導系の構築; ホニュウルイ ランホウ ケイセイ キコウ ノ カイセキ ： ランホウ ハツイク ニ オケル ギャップ ケツゴウ ノ カイセキ ト マウス ハイセイカン サイボウ カラノ ランホウ ユウドウケイ ノ コウチク. (Thesis). Nara Institute of Science and Technology / 奈良先端科学技術大学院大学. Retrieved from http://hdl.handle.net/10061/5726
Note: this citation may be lacking information needed for this citation format:
No year of publication.
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Nitta, Mai. “Approaches to understanding the mechanism of mammalian folliculogenesis : Analysis of gap junctions in ovarian follicles, and the establishment of an in vitro system for obtaining follicles from mouse embryonic stem cells : 哺乳類卵胞形成機構の解析：卵胞発育におけるギャップ結合の解析とマウス胚性幹細胞からの卵胞誘導系の構築; ホニュウルイ ランホウ ケイセイ キコウ ノ カイセキ ： ランホウ ハツイク ニ オケル ギャップ ケツゴウ ノ カイセキ ト マウス ハイセイカン サイボウ カラノ ランホウ ユウドウケイ ノ コウチク.” Thesis, Nara Institute of Science and Technology / 奈良先端科学技術大学院大学. Accessed September 17, 2019.
http://hdl.handle.net/10061/5726.
Note: this citation may be lacking information needed for this citation format:
No year of publication.
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Nitta, Mai. “Approaches to understanding the mechanism of mammalian folliculogenesis : Analysis of gap junctions in ovarian follicles, and the establishment of an in vitro system for obtaining follicles from mouse embryonic stem cells : 哺乳類卵胞形成機構の解析：卵胞発育におけるギャップ結合の解析とマウス胚性幹細胞からの卵胞誘導系の構築; ホニュウルイ ランホウ ケイセイ キコウ ノ カイセキ ： ランホウ ハツイク ニ オケル ギャップ ケツゴウ ノ カイセキ ト マウス ハイセイカン サイボウ カラノ ランホウ ユウドウケイ ノ コウチク.” Web. 17 Sep 2019.
Note: this citation may be lacking information needed for this citation format:
No year of publication.
Vancouver:
Nitta M. Approaches to understanding the mechanism of mammalian folliculogenesis : Analysis of gap junctions in ovarian follicles, and the establishment of an in vitro system for obtaining follicles from mouse embryonic stem cells : 哺乳類卵胞形成機構の解析：卵胞発育におけるギャップ結合の解析とマウス胚性幹細胞からの卵胞誘導系の構築; ホニュウルイ ランホウ ケイセイ キコウ ノ カイセキ ： ランホウ ハツイク ニ オケル ギャップ ケツゴウ ノ カイセキ ト マウス ハイセイカン サイボウ カラノ ランホウ ユウドウケイ ノ コウチク. [Internet] [Thesis]. Nara Institute of Science and Technology / 奈良先端科学技術大学院大学; [cited 2019 Sep 17].
Available from: http://hdl.handle.net/10061/5726.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
No year of publication.
Council of Science Editors:
Nitta M. Approaches to understanding the mechanism of mammalian folliculogenesis : Analysis of gap junctions in ovarian follicles, and the establishment of an in vitro system for obtaining follicles from mouse embryonic stem cells : 哺乳類卵胞形成機構の解析：卵胞発育におけるギャップ結合の解析とマウス胚性幹細胞からの卵胞誘導系の構築; ホニュウルイ ランホウ ケイセイ キコウ ノ カイセキ ： ランホウ ハツイク ニ オケル ギャップ ケツゴウ ノ カイセキ ト マウス ハイセイカン サイボウ カラノ ランホウ ユウドウケイ ノ コウチク. [Thesis]. Nara Institute of Science and Technology / 奈良先端科学技術大学院大学; Available from: http://hdl.handle.net/10061/5726
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
No year of publication.
University of Edinburgh
2.
Meek, Stephen Earl.
Experimental approaches to establish rat embryonic stem cells.
Degree: PhD, 2011, University of Edinburgh
URL: http://hdl.handle.net/1842/5040
► The rat has been an established experimental animal model within many areas of biological investigation for over one hundred years due to its size, breeding…
(more)
▼ The rat has been an established experimental animal model within many areas of biological investigation for over one hundred years due to its size, breeding characteristics, and knowledge of its physiology and behaviour. In recent years its status as a leading biomedical model has been somewhat surpassed by the mouse. This is largely the result of the isolation and application of mouse embryonic stem (ES) cells. Mouse ES cells have the capacity for unlimited self-renew in vitro whilst maintaining pluripotency and germline competence, and most importantly are amenable to sophisticated reverse genetics strategies such as gene targeting, which have provided a route to germ line modification. Thus far, the derivation of rat ES cells has proved elusive. The generation of rat ES cells would therefore facilitate equivalent applications to rat genetics and significantly strengthen the rat as an experimental model system. Previous attempts to derive rat ES cells led to the isolation of rat ES-like cells. However, whilst these cells exhibit extensive self-renew in vitro, it was known that they fail to maintain significant levels of the key functional ES cell marker Oct4 and do not contribute to chimeras. Rather, these cells express the trophectoderm markers Cdx2 and CyclinD3, and have been termed ExS cells due to their probable extra-embryonic nature. In the work described in this thesis, further investigation of ExS cells revealed the absence of expression of the key pluripotency gene Nanog, although the expression pattern of Nanog in the rat embryo was shown to be similar to that of mouse. It was hypothesised that expression of exogenous Oct4 and Nanog or Sox2 genes could facilitate reprogramming of ExS cells into a 'true' ES cell state. Initial work described in this thesis demonstrated that it was possible to introduce transgenes into rat ExS cells and obtain stable transformants with long term transgene expression. On this basis Oct 4, Nanog and Sox2 transgene expression vectors were constructed and stably integrated into ExS cells, and transgene expression verified. However, no reactivation of an endogenous gene expression profile, characteristic of a true ES cell-like state, was observed in any of the transgenic lines produced. Concurrent with work on ExS cells, investigations by others using chemically defined, serum-free medium containing small molecule inhibitors of MEK and GSK3 (called 3i/2i medium) had demonstrated that it was possible to readily isolate mouse ES cells, even from strains known to be refractory to ES cell isolation. Therefore, the ability of this culture system to facilitate rat ES cell derivation was investigated. Rat 3i/2i cell lines were established from ICM outgrowths of Fischer, DA and Sprague Dawley E4.5 rat embryos. These cells maintained expression of Oct4 and Nanog and could generate complex teratomas consisting of all three germ layers. They were distinct from epiblast stem cells (EpiSC) in that they expressed Klf4, Rex1 and Stella and most importantly, they could contribute to the…
Subjects/Keywords: 571.6; ES cells; 2i; rat; Oct4; Nanog
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APA (6th Edition):
Meek, S. E. (2011). Experimental approaches to establish rat embryonic stem cells. (Doctoral Dissertation). University of Edinburgh. Retrieved from http://hdl.handle.net/1842/5040
Chicago Manual of Style (16th Edition):
Meek, Stephen Earl. “Experimental approaches to establish rat embryonic stem cells.” 2011. Doctoral Dissertation, University of Edinburgh. Accessed September 17, 2019.
http://hdl.handle.net/1842/5040.
MLA Handbook (7th Edition):
Meek, Stephen Earl. “Experimental approaches to establish rat embryonic stem cells.” 2011. Web. 17 Sep 2019.
Vancouver:
Meek SE. Experimental approaches to establish rat embryonic stem cells. [Internet] [Doctoral dissertation]. University of Edinburgh; 2011. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/1842/5040.
Council of Science Editors:
Meek SE. Experimental approaches to establish rat embryonic stem cells. [Doctoral Dissertation]. University of Edinburgh; 2011. Available from: http://hdl.handle.net/1842/5040
University of Edinburgh
3.
Wachter, Elisabeth.
Influence of CpG islands on chromatin structure.
Degree: PhD, 2014, University of Edinburgh
URL: http://hdl.handle.net/1842/9369
► CpG islands (CGIs) are short GC rich sequences with a high frequency of CpGs that are associated with the active chromatin mark H3K4me3. Most occur…
(more)
▼ CpG islands (CGIs) are short GC rich sequences with a high frequency of CpGs that are associated with the active chromatin mark H3K4me3. Most occur at gene promoters and are often free of cytosine methylation. Recent work has begun to clarify the functional significance of CGIs with respect to chromatin structure and transcription. In particular, proteins associated with histone-modifying activities, such as Cfp1 and Kdm2a, bind specifically to non-methylated CGIs via their CxxC domains. For example, artificial promoterless CpG-rich sequences integrated at the 3’ UTR of genes recruit Cfp1 and generate novel peaks of H3K4me3 in mouse ES cells without apparent RNA polymerase recruitment. There is also evidence that G+C-rich DNA recruits H3K27me3, a gene silencing mark. In this thesis I am exploring the constraints on DNA sequence and genomic location that are required to impose both H3K4me3 and H3K27me3 at CGI sequences. Showing that the generation of novel peaks of H3K4me3 and H3K27me3 over a promoter-less CpG rich sequence in a gene desert region is independent of it’s location in the genome extends earlier findings. These findings suggest that shared features of the primary DNA sequence at CGIs directly influence chromatin modification. Thus CGIs are not passive footprints of other cellular mechanisms, but play an active role in setting up local chromatin structure. However, the relative contribution of CpG frequency versus G+C content remains unclear. Therefore a sequence was generated that contains low levels of CpGs, comparable to the bulk genome, but has a G+C content similar to that of CGIs (Low CpG / High G+C). When this sequence was inserted into a gene desert neither marks of H3K4me3 or H3K27me3 were formed, indicating the importance of CpGs. Surprisingly, the reverse sequence with a high CpG frequency similar to that of CGIs and a low G+C content similar to that of the bulk genome (High CpG / Low G+C) did not establish H3K4me3 or H3K27me3 either. However, it was found that this sequence becomes heavily methylated in contrast to CGI-like sequences that remained unmethylated when introduced into a gene desert. This finding suggests that a high G+C content is important for keeping CGI-like sequences methylation free. Upon insertion of this High CpG / Low G+C sequence into mouse ES cells that were devoid of the de-novo DNA methyltransferases 3a and 3b (Dnmt3a/3b -/-) both H3K4me3 and H3K27me3 marks were established at the inserted sequence. This discovery confirms the importance of CpGs for setting up local chromatin structure.
Subjects/Keywords: 572.8; CpG islands; chromatin; polycomb; ES cells
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APA ·
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APA (6th Edition):
Wachter, E. (2014). Influence of CpG islands on chromatin structure. (Doctoral Dissertation). University of Edinburgh. Retrieved from http://hdl.handle.net/1842/9369
Chicago Manual of Style (16th Edition):
Wachter, Elisabeth. “Influence of CpG islands on chromatin structure.” 2014. Doctoral Dissertation, University of Edinburgh. Accessed September 17, 2019.
http://hdl.handle.net/1842/9369.
MLA Handbook (7th Edition):
Wachter, Elisabeth. “Influence of CpG islands on chromatin structure.” 2014. Web. 17 Sep 2019.
Vancouver:
Wachter E. Influence of CpG islands on chromatin structure. [Internet] [Doctoral dissertation]. University of Edinburgh; 2014. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/1842/9369.
Council of Science Editors:
Wachter E. Influence of CpG islands on chromatin structure. [Doctoral Dissertation]. University of Edinburgh; 2014. Available from: http://hdl.handle.net/1842/9369
University of Vienna
4.
Abdeen, Dana.
Identification of novel regulators of sprouting angiogenesis using haploid ES cells.
Degree: 2017, University of Vienna
URL: http://othes.univie.ac.at/50257/
► Die Expansion, Remodellierung und Reifung von Blutgefäßen ist ein wichtiges physiologisches Ereignis, das die embryonalen Entwicklungsstadien erfüllt und an der Pathophysiologie der Erwachsenen wie Augenerkrankungen…
(more)
▼ Die Expansion, Remodellierung und Reifung von Blutgefäßen ist ein wichtiges physiologisches Ereignis, das die embryonalen Entwicklungsstadien erfüllt und an der Pathophysiologie der Erwachsenen wie Augenerkrankungen oder Krebsentwicklung beteiligt ist. Die Kaskade von Ereignissen, die zur Bildung neuer Blutgefäße aus bereits bestehenden führt, wird als Angiogenese bezeichnet. Viele bekannte und doch wenig verstandene Signalwege koordinieren und verfeinern den angiogenen Prozess. In diesem Zusammenhang sind genetische Funktionsverlustbildschirme ein wirkungsvolles Werkzeug, um die Rolle möglicher Spieler aufzudecken.
Unser Ziel war es, neue angiogene Regulatoren zu finden, indem wir In-vitro-Keimungsmodelle entwickeln, die die Angiogenese von Sprossen in vivo nachahmen. Wir haben dementsprechend 32 Kandidatengene ausgewählt und eine vergleichende Studie durchgeführt, die Wildtyp- und Mutanten-Genfunktionen umfasst. Der Vergleich basierte auf der Aufklärung der Auswirkungen von Genmutationen auf die sprießenden Phänotypen in 3D-Kollagengelen. Zu diesem Zweck verwendeten wir bedingt mutagenisierte haploide murine ES-Zellen. Für jedes erhaltene Kandidatengen erzeugten wir Schwesterklone, die sich nur darin unterschieden, das Gen von Interesse entweder in der mutierten oder in der Wildtypform unter Verwendung von Cre-Rekombinationstechnologien zu tragen. Wir führten auch Kompetitionsstudien durch, indem wir gleichartige Mischungen von Wildtyp- und Mutantenklonen subkutan in immunsupprimierte Mäuse injizierten und ihren differentiellen Beitrag zur Teratomvaskulatur mittels Fluoreszenz-aktivierter Zellsortierung (FACS) -Analyse und Immunhistochemie validierten.
Zusammenfassend zeigten unsere Ergebnisse, dass die Mutagenese von 13 Kandidatengenen zu einer signifikanten Abweichung ihrer sprossenden Phänotypen im Vergleich zu ihren Wildtypschwestern führte, die entweder eine verminderte oder eine erhöhte Keimfähigkeit aufwiesen. Wichtig ist, dass der sprießende Phänotyp unserer Top-Hits in In-vivo-Experimenten bestätigt wurde. Daher haben wir die Funktion von mehreren neuartigen Angiogenese-Genen identifiziert und validiert.
Blood vessel expansion, remodelling and maturation is a major physiological event that fulfils the embryonic developmental stages and takes part in adult pathophysiology such as ocular diseases or cancer development. The cascade of events that leads to the formation of new blood vessels from already existing ones is termed angiogenesis. Many known and yet poorly understood pathways orchestrate and fine tune the angiogenic process. In this context, genetic loss-of-function screens are a powerful tool to uncover the role of possible players.
Our goal was to find novel angiogenic regulators by designing in vitro sprouting models that mimic sprouting angiogenesis in vivo. We accordingly chose 32 candidate genes and performed a comparative study comprising wildtype and a mutant gene functions. The comparison was based on elucidating the effect of gene mutations on the sprouting phenotypes in 3D collagen…
Subjects/Keywords: 42.13 Molekularbiologie; Angiogenesis / Haploid ES cells / Hyposprouters / Hypersprouters; Angiogenesis / Haploid ES cells / Hyposprouters / Hypersprouters
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APA (6th Edition):
Abdeen, D. (2017). Identification of novel regulators of sprouting angiogenesis using haploid ES cells. (Thesis). University of Vienna. Retrieved from http://othes.univie.ac.at/50257/
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Abdeen, Dana. “Identification of novel regulators of sprouting angiogenesis using haploid ES cells.” 2017. Thesis, University of Vienna. Accessed September 17, 2019.
http://othes.univie.ac.at/50257/.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Abdeen, Dana. “Identification of novel regulators of sprouting angiogenesis using haploid ES cells.” 2017. Web. 17 Sep 2019.
Vancouver:
Abdeen D. Identification of novel regulators of sprouting angiogenesis using haploid ES cells. [Internet] [Thesis]. University of Vienna; 2017. [cited 2019 Sep 17].
Available from: http://othes.univie.ac.at/50257/.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Abdeen D. Identification of novel regulators of sprouting angiogenesis using haploid ES cells. [Thesis]. University of Vienna; 2017. Available from: http://othes.univie.ac.at/50257/
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
University of Edinburgh
5.
Cowan, Gillian.
Fetal germ cell differentiation and the impact of the somatic cells.
Degree: 2009, University of Edinburgh
URL: http://hdl.handle.net/1842/4164
► Specification of a germ cell lineage and appropriate maturation are essential for the transfer of genetic information from one generation to the next. Germ cells…
(more)
▼ Specification of a germ cell lineage and appropriate maturation are essential for the transfer of genetic information from one generation to the next. Germ cells form from pluripotent precursor cells that migrate into the gonadal ridge and undergo commitment to either the female or male lineage. In the fetal ovary, germ cells enter meiotic prophase I, then arrest at the diplotene stage; in the testis germ cells do not begin meiosis until puberty. Abnormal differentiation of germ cells can result in malignant transformation. Somatic cells play a key role in modulating the developmental fate of the germ cells. Research into germ cell development during fetal life has almost exclusively focused on studies in rodents, but we, and others, have reported several fundamental differences in the expression of germ cell specific markers in the human compared with the mouse. The studies described in this thesis have investigated germ cell-specific gene expression and the possible impact of the somatic cells during development. This was achieved by studying human fetal gonads obtained during the 1st and 2nd trimesters of pregnancy and through the use of both wild-type and mutant mouse ES cell lines. Studies on germ cells in the human fetal testis have extended the findings of others, and confirmed that germ cell populations at different stages of maturation co-exist in the human fetal testis, a situation that is in contrast to that in rodents. For example expression of M2A and AP2γ was restricted to the OCT4-positive gonocyte population, while VASA and NANOS1 were localised exclusively to the to the OCT4-negative prespermatogonia. DAZL was expressed in both populations. Analysis also revealed that both the gonocyte and prespermatogonial populations proliferate throughout the 2nd trimester. Recent studies have implicated retinoic acid (RA) in the control of meiotic entry in germ cells of the fetal mouse ovary. In this study we demonstrated for the first time that two genes implicated in the action of RA in mouse gonad, STRA8 and NANOS2, are also expressed in a similar sexspecific- manner in the human fetal gonads, and that the RA receptors are present in both somatic and germ cells suggesting that RA may regulate germ cell function in the human as well as the mouse. However, whilst the mesonephros appears to be the primary site of RA synthesis in the mouse our initial studies indicate that in the human the gonad itself may be a more likely site of RA biosynthesis. In the fetal mouse testis, RA is degraded by the enzyme Cyp26b1 present in the somatic cells and germ cells do not enter meiosis, our novel findings suggest that CYP26B1 is more abundant in the human fetal ovary than the testis, suggesting that meiotic entry may be controlled by an alternative signalling pathway in the human. One of the methods that can aid our understanding of somatic cell gene expression in the gonad is in vitro culture. To date, there have been no published reports of the successful in vitro culture of somatic cells from the human fetal testis. In the…
Subjects/Keywords: 612.6; germ cells; sertoli cells; ES cells; meiosis
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APA ·
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Export
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APA (6th Edition):
Cowan, G. (2009). Fetal germ cell differentiation and the impact of the somatic cells. (Doctoral Dissertation). University of Edinburgh. Retrieved from http://hdl.handle.net/1842/4164
Chicago Manual of Style (16th Edition):
Cowan, Gillian. “Fetal germ cell differentiation and the impact of the somatic cells.” 2009. Doctoral Dissertation, University of Edinburgh. Accessed September 17, 2019.
http://hdl.handle.net/1842/4164.
MLA Handbook (7th Edition):
Cowan, Gillian. “Fetal germ cell differentiation and the impact of the somatic cells.” 2009. Web. 17 Sep 2019.
Vancouver:
Cowan G. Fetal germ cell differentiation and the impact of the somatic cells. [Internet] [Doctoral dissertation]. University of Edinburgh; 2009. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/1842/4164.
Council of Science Editors:
Cowan G. Fetal germ cell differentiation and the impact of the somatic cells. [Doctoral Dissertation]. University of Edinburgh; 2009. Available from: http://hdl.handle.net/1842/4164
University of Edinburgh
6.
Jin, Xin.
Towards differentiation of mouse embryonic stem cells to thymic epithelial progenitor cells.
Degree: PhD, 2013, University of Edinburgh
URL: http://hdl.handle.net/1842/12227
► The thymus is the major site for T-cell generation and thus is important for the adaptive immune system. Development of a properly selected, functional T-cell…
(more)
▼ The thymus is the major site for T-cell generation and thus is important for the adaptive immune system. Development of a properly selected, functional T-cell repertoire relies on interactions between developing T cells and a series of functionally distinct thymic stroma cell types including the cortical and medullary thymic epithelial cells (TECs). The thymus is one of the first organs to degenerate in normal healthy ageing. Related to this, there is strong interest in developing protocols for improving thymus function in patients by cell replacement or regenerative therapies. Thymic epithelial progenitor cells (TEPCs) represent a potential source of cells for thymus transplantation. However, the only source of these cells for transplantation is currently fetal thymus tissue. If TEPCs could be generated from pluripotent cells, this could provide an alternative source of cells for transplantation. The work described in this thesis therefore had two central aims (i) to test the stability of thymic epithelial progenitor cells in vivo and (ii) to investigate the possibility of generating TEPCs or TECs from mouse embryonic stem (ES) cells. The forkhead transcription factor, Foxn1, is essential for the development of a functionally mature thymic epithelium, but is not necessary for formation of the thymic primordium or for medullary thymic epithelial sub-lineage specification. By reactivating Foxn1 expression postnatally in mice carrying a revertible hypomorphic allele of Foxn1, Foxn1R, I herein demonstrate that TEPCs that can express only low levels of Foxn1 mRNA can persist postnatally in the thymic rudiment in mice until at least 6 months of age, and retain the potential to give rise to both cortical and medullary thymic epithelial cells (cTECs and mTECs). These data demonstrate that the TEPC-state is remarkably stable in vivo under conditions of low Foxn1 expression. In parallel with this work, I confirmed the possibility of generating Foxn1-expressing cells from mouse ES cells by using a Foxn1 reporter cell line. As the thymic epithelium has a single origin in the third pharyngeal pouch (3pp) endoderm, I then tested whether or not TEPCs and /or TECs were generated during ES cell differentiation via existing protocols for generating anterior definitive endoderm differentiation cells from mouse ES cells. From this work, I showed that genes expressed in the 3pp and/or TEPC,-including Plet-1, Tbx1, Hoxa3 and Pax9, were induced by differentiation of ES cells using these protocols. I further showed that cells expressing both Plet-1, a marker of foregut endoderm and 3pp, and EpCAM, a marker of proliferating epithelial cells, were induced using a novel protocol (2i ADE) for generating ES cells from ADE. However, gene expression analysis and functional testing suggested that the majority of these cells were non-thymus lineage. I subsequently developed a novel protocol which combined this 2i ADE protocol with co-culturing of the differentiating ES cells with fetal thymic lobes, and demonstrated that this further induced 3pp…
Subjects/Keywords: 611; thymus; TEPC; thymic epithelial cells; embryonic stem cells; ES cells
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APA (6th Edition):
Jin, X. (2013). Towards differentiation of mouse embryonic stem cells to thymic epithelial progenitor cells. (Doctoral Dissertation). University of Edinburgh. Retrieved from http://hdl.handle.net/1842/12227
Chicago Manual of Style (16th Edition):
Jin, Xin. “Towards differentiation of mouse embryonic stem cells to thymic epithelial progenitor cells.” 2013. Doctoral Dissertation, University of Edinburgh. Accessed September 17, 2019.
http://hdl.handle.net/1842/12227.
MLA Handbook (7th Edition):
Jin, Xin. “Towards differentiation of mouse embryonic stem cells to thymic epithelial progenitor cells.” 2013. Web. 17 Sep 2019.
Vancouver:
Jin X. Towards differentiation of mouse embryonic stem cells to thymic epithelial progenitor cells. [Internet] [Doctoral dissertation]. University of Edinburgh; 2013. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/1842/12227.
Council of Science Editors:
Jin X. Towards differentiation of mouse embryonic stem cells to thymic epithelial progenitor cells. [Doctoral Dissertation]. University of Edinburgh; 2013. Available from: http://hdl.handle.net/1842/12227
University of Toronto
7.
Vexelshtein, Olga.
DNA Methylation of Endothelial Cell-enriched Genes during in Vitro Mouse ES Cell Differentiation and the Role of TET Proteins.
Degree: 2016, University of Toronto
URL: http://hdl.handle.net/1807/72845
► Endothelial cell (EC)-enriched genes are regulated by numerous mechanisms, but especially epigenetic mechanisms such as DNA methylation. The current work examines the importance of TET1…
(more)
▼ Endothelial cell (EC)-enriched genes are regulated by numerous mechanisms, but especially epigenetic mechanisms such as DNA methylation. The current work examines the importance of TET1 and 2 in murine embryonic stem (ES) cells and their relevance to EC-enriched gene expression. The proximal promoters of EC-enriched genes are differentially methylated in expressing versus non-expressing cell types. To examine how this methylation pattern between EC and non-EC is established, we studied ES cells. The DNA methylation pattern was heterogeneous, and partially propagated to daughter cells upon cell division. When in vitro differentiating WT and TET1 and 2 double knockout (DKO) ES cells, the DNA methylation of EC-enriched gene promoters increases at day 4 and day 7 in both cell lines. Despite their hypermenthylation at day 7, EC- enriched gene expression increases in the WT cells but it is blunted in the DKO cells. Suggesting that TET1 and 2 presence in ES cells facilitate the activation of EC-enriched genes later in development.
M.Sc.
Advisors/Committee Members: Marsden, Philip A, Medical Biophysics.
Subjects/Keywords: DNA methylation; Endothelial cells; ES cells; TET proteins; 0379
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APA (6th Edition):
Vexelshtein, O. (2016). DNA Methylation of Endothelial Cell-enriched Genes during in Vitro Mouse ES Cell Differentiation and the Role of TET Proteins. (Masters Thesis). University of Toronto. Retrieved from http://hdl.handle.net/1807/72845
Chicago Manual of Style (16th Edition):
Vexelshtein, Olga. “DNA Methylation of Endothelial Cell-enriched Genes during in Vitro Mouse ES Cell Differentiation and the Role of TET Proteins.” 2016. Masters Thesis, University of Toronto. Accessed September 17, 2019.
http://hdl.handle.net/1807/72845.
MLA Handbook (7th Edition):
Vexelshtein, Olga. “DNA Methylation of Endothelial Cell-enriched Genes during in Vitro Mouse ES Cell Differentiation and the Role of TET Proteins.” 2016. Web. 17 Sep 2019.
Vancouver:
Vexelshtein O. DNA Methylation of Endothelial Cell-enriched Genes during in Vitro Mouse ES Cell Differentiation and the Role of TET Proteins. [Internet] [Masters thesis]. University of Toronto; 2016. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/1807/72845.
Council of Science Editors:
Vexelshtein O. DNA Methylation of Endothelial Cell-enriched Genes during in Vitro Mouse ES Cell Differentiation and the Role of TET Proteins. [Masters Thesis]. University of Toronto; 2016. Available from: http://hdl.handle.net/1807/72845
Texas A&M University
8.
Harding, Heather Darby.
Oct-4 expression in equine embryonic cells.
Degree: 2007, Texas A&M University
URL: http://hdl.handle.net/1969.1/4793
► The Oct-4 transcription factor is believed to co-regulate early embryonic development of mammals due to the correlation of its presence with the maintenance of pluripotency.…
(more)
▼ The Oct-4 transcription factor is believed to co-regulate early embryonic
development of mammals due to the correlation of its presence with the maintenance of
pluripotency. It is commonly used as a marker for the identification of embryonic stem
(
ES)
cells for this reason. Until 1999, Oct-4 studies were limited to in vivo-produced
embryos; equine embryos have not been studied for their Oct-4 expression patterns. In
addition, equine stem-like
cells (defined by marker expression, induced differentiation,
passage survival, and morphology) have recently been isolated from in vivo-produced
embryos, but no work has been performed in horses to isolate
ES cells from in vitroproduced
embryos.
This study investigated the expression of Oct-4 transcription factor using
immunocytochemistry in 42 in vitro-produced embryos aged 1-10 days and in 5 in vivoproduced
blastocysts aged 7-10 days. Effective conditions for rapid establishment of a
feeder layer of equine fetal fibroblasts were established, and this feeder layer was used to
grow isolated equine inner cell mass (ICM)
cells from in vitro-produced embryos. The
expression of Oct-4 was examined in resultant cell growths.
In vitro-produced embryos less than 6 days of age showed variable staining
within blastomeres of the same embryo, and the peak of variability correlated with maternal-zygotic transition. After Oct-4 staining of in vitro-produced blastocysts, no
cells could be identified as an ICM based on a difference in fluorescent intensity from
the other
cells of the blasyocysts. However, in vitro-produced blastocysts that were
subsequently cultured in vivo contained a presumptive ICM, visible based on greater
fluorescent intensity of Oct-4 stain. The trophoblast of all blastocysts also stained
positively for Oct-4 protein. Fibroblasts were successfully isolated from equine feti.
Treatment with 20 ????g/ml of Mitomycin C arrested cell growth without causing excessive
death. Fibroblasts were inactivated and frozen, then thawed as needed to establish a
confluent monolayer for ICM isolation overnight. ICMs from in vitro-produced
embryos formed outgrowths, but none that could be identified morphologically as
ES
cells. Outgrowth
cells contained about 20% Oct-4 expressing
cells in sporadic
groupings. Assuming appropriate binding of the Oct-4 antibody, Oct-4 expressing
cells
(potentially indicating pluripotency) are found throughout the embryo in early
development and in the feeder layer after co-culture.
Advisors/Committee Members: Hinrichs, Katrin (advisor), Binas, Bert (committee member), Choi, Young Ho (committee member), Westhusin, Mark (committee member).
Subjects/Keywords: Oct-4; ES cells; pluripotency
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APA (6th Edition):
Harding, H. D. (2007). Oct-4 expression in equine embryonic cells. (Thesis). Texas A&M University. Retrieved from http://hdl.handle.net/1969.1/4793
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Harding, Heather Darby. “Oct-4 expression in equine embryonic cells.” 2007. Thesis, Texas A&M University. Accessed September 17, 2019.
http://hdl.handle.net/1969.1/4793.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Harding, Heather Darby. “Oct-4 expression in equine embryonic cells.” 2007. Web. 17 Sep 2019.
Vancouver:
Harding HD. Oct-4 expression in equine embryonic cells. [Internet] [Thesis]. Texas A&M University; 2007. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/1969.1/4793.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Harding HD. Oct-4 expression in equine embryonic cells. [Thesis]. Texas A&M University; 2007. Available from: http://hdl.handle.net/1969.1/4793
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Case Western Reserve University
9.
Schnetz, Michael Paul.
Investigation of the Molecular Function of CHD7, the Protein
Implicated in CHARGE Syndrome, Using Next-Generation
Genomics.
Degree: PhD, Genetics, 2010, Case Western Reserve University
URL: http://rave.ohiolink.edu/etdc/view?acc_num=case1275581022
► Eukaryotes use chromatin to package large, complex genomes into the nucleus of their cells. As such, chromatin structure plays a key role in virtually all…
(more)
▼ Eukaryotes use chromatin to package large, complex
genomes into the nucleus of their
cells. As such, chromatin
structure plays a key role in virtually all DNA-centric processes
including gene transcription. Specifically, chromatin packaging and
specific histone chemical modifications function to regulate gene
expression. The chromodomain helicase DNA-binding (CHD) protein
family is a class of chromatin remodeling enzymes that play roles
in these processes. Despite what is known about the CHD protein
family, the molecular function of CHD7 has not been characterized.
CHD7 is required for proper mammalian development as mutations in
CHD7 result in a developmental disease known as CHARGE syndrome in
which multiple organ systems are affected including the eyes,
heart, and genitals. Using a genomics approach, the molecular
function of CHD7 was investigated. CHD7 was found to bind chromatin
largely outside of gene transcriptional start sites in an overall
pattern that was cell-specific. CHD7 binding correlates with
cell-specific gene expression and harbors features of gene enhancer
elements, namely H3K4me1 and DNaseI hypersensitivity. CHD7, to a
less extent, is found also at gene promoters marked with H3K4me3.
CHD7 binds H3K4me via its chromodomains. In mouse
ES cells, CHD7
binding tracks with H3K4me1 methylation during cellular
differentiation and colocalizes with
ES-specific transcription
factors including Oct4, Nanog, and Sox2. CHD7 binding sites
represent functional enhancers as tested in luciferase reporter
assays. Finally, loss of CHD7 results in both expression increases
as well as decreases, however, CHD7 binding correlates with
increased gene expression. Collectively, these studies indicate
that CHD7 functions to regulate cell-specific gene transcription
through enhancer interactions by limiting the expression of its
target genes. The function of CHD7 uncovers a novel regulatory
mechanism at gene enhancers that suggests the overall regulatory
output of any enhancer is the combination of both positive and
negative-acting factors.
Advisors/Committee Members: Scacheri, Peter (Advisor).
Subjects/Keywords: Genetics; CHD7; CHARGE Syndrome; Chromatin Remodeling; mouse ES cells
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APA (6th Edition):
Schnetz, M. P. (2010). Investigation of the Molecular Function of CHD7, the Protein
Implicated in CHARGE Syndrome, Using Next-Generation
Genomics. (Doctoral Dissertation). Case Western Reserve University. Retrieved from http://rave.ohiolink.edu/etdc/view?acc_num=case1275581022
Chicago Manual of Style (16th Edition):
Schnetz, Michael Paul. “Investigation of the Molecular Function of CHD7, the Protein
Implicated in CHARGE Syndrome, Using Next-Generation
Genomics.” 2010. Doctoral Dissertation, Case Western Reserve University. Accessed September 17, 2019.
http://rave.ohiolink.edu/etdc/view?acc_num=case1275581022.
MLA Handbook (7th Edition):
Schnetz, Michael Paul. “Investigation of the Molecular Function of CHD7, the Protein
Implicated in CHARGE Syndrome, Using Next-Generation
Genomics.” 2010. Web. 17 Sep 2019.
Vancouver:
Schnetz MP. Investigation of the Molecular Function of CHD7, the Protein
Implicated in CHARGE Syndrome, Using Next-Generation
Genomics. [Internet] [Doctoral dissertation]. Case Western Reserve University; 2010. [cited 2019 Sep 17].
Available from: http://rave.ohiolink.edu/etdc/view?acc_num=case1275581022.
Council of Science Editors:
Schnetz MP. Investigation of the Molecular Function of CHD7, the Protein
Implicated in CHARGE Syndrome, Using Next-Generation
Genomics. [Doctoral Dissertation]. Case Western Reserve University; 2010. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=case1275581022
University of Melbourne
10.
Motazedian, Ali.
Lymphocyte differentiation from pluripotent stem cells.
Degree: 2017, University of Melbourne
URL: http://hdl.handle.net/11343/191658
► Human pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), are cell types that can undergo unlimited self-renewal and,…
(more)
▼ Human pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), are cell types that can undergo unlimited self-renewal and, in theory, have the ability to give rise to all of the different cell types in the body. These properties of PSCs make them an attractive platform for generating a variety of normal and mutated human cell types for medical related applications.
In this study, we investigated methods for generating B-cells and T-cells from PSCs. B and T lymphocytes together comprise the adaptive immune system. One of the functions of T-cells is to find and destroy defective cells, a function that has been co-opted into promising cell based therapies for the treatment of CD19+ leukaemia. Similarly, antibodies produced by B-cells now underlie promising treatments for a range of malignancies, including melanoma and lung cancer. Moreover, B-lymphocytes are of particular interest as their derivation would be indicative of progress towards the development of conditions that support hematopoietic stem cells (HSC) formation, the cell type which is likely to have the most immediate clinical applications. Therefore, in vitro production of both T and B cells from PSCs could facilitate the development of a new generation of therapeutics derived from these cell types. In this project, we examined both transcription factor and growth factor mediated induction of B-lymphoid cells as well as culture conditions that supported “stromal free” T-cell development.
First, starting with culture conditions that supported primitive hematopoietic differentiation of PSCs, we investigated the effect of over-expressing B-lineage specific transcription factors in an inducible manner. Our experiments showed that, during the emergence of primitive blood cells, forced expression of PAX5 alone in CD34+ hematopoietic progenitors up-regulated the B-cell marker CD19 and, paradoxically, promoted the appearance of Glycophorin A expressing erythroid lineage cells. However, CD10+CD19+ B-cell progenitors were not observed, indicating that in the context of the primitive blood development, CD34+ progenitors were not competent to execute the instructions carried by PAX5.
Second, we investigated the lymphogenic characteristics of AGM-like structures developed during a definitive hematopoiesis differentiation protocol established in laboratory. We showed that, T-lineage specification could occur in the absence of exogenous Notch1 ligands and without the need for purification of CD34+ progenitors by FACS. This finding will pave the way for a scalable method for generating T-cells for both research and for potential clinical applications.
Lastly, having seen how AGM-like structures facilitated T-cell differentiation, we revisited our initial goal of B-lymphocyte differentiation from PSCs. This time, using growth factor directed differentiation, we were able to demonstrate reproducible generation of B-lymphoid progenitors from multiple PSC lines.
Taken together, we hope our findings will aid the…
Subjects/Keywords: pluripotent stem cells; iPS; ES; lymphocyte; T-cell; B-cell
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APA (6th Edition):
Motazedian, A. (2017). Lymphocyte differentiation from pluripotent stem cells. (Doctoral Dissertation). University of Melbourne. Retrieved from http://hdl.handle.net/11343/191658
Chicago Manual of Style (16th Edition):
Motazedian, Ali. “Lymphocyte differentiation from pluripotent stem cells.” 2017. Doctoral Dissertation, University of Melbourne. Accessed September 17, 2019.
http://hdl.handle.net/11343/191658.
MLA Handbook (7th Edition):
Motazedian, Ali. “Lymphocyte differentiation from pluripotent stem cells.” 2017. Web. 17 Sep 2019.
Vancouver:
Motazedian A. Lymphocyte differentiation from pluripotent stem cells. [Internet] [Doctoral dissertation]. University of Melbourne; 2017. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/11343/191658.
Council of Science Editors:
Motazedian A. Lymphocyte differentiation from pluripotent stem cells. [Doctoral Dissertation]. University of Melbourne; 2017. Available from: http://hdl.handle.net/11343/191658
11.
Osorno Hernandez, Carlos Rodrigo.
Transcription factor heterogeneity in epiblast pluripotency.
Degree: PhD, 2013, University of Edinburgh
URL: http://hdl.handle.net/1842/8081
► Pluripotency is the ability of a cell to differentiate into derivatives of all three somatic lineages and germ cells. In vivo, pluripotent cells exist transiently…
(more)
▼ Pluripotency is the ability of a cell to differentiate into derivatives of all three somatic lineages and germ cells. In vivo, pluripotent cells exist transiently in the epiblast of the developing embryo and in rare tumour cells. In vitro, pluripotent cells have been isolated and propagated from teratocarcinomas (EC cells), preimplantation epiblast (ES cells) and post-implantation epiblast (EpiSCs). Pluripotency is governed by a gene regulatory network centred on the triumvirate of transcription factors Oct4, Sox2 and Nanog. Interestingly, transcription factors that are important to direct pluripotent cell identity are not all equally distributed throughout the pluripotent cell population. While Oct4 levels are relatively homogeneous, other transcription factors, such as Nanog, are more heterogeneously expressed. Additionally, an increasing body of evidence indicates that extrinsic cues also play a critical role in the establishment and maintenance of pluripotency. Using biochemical and genetic tools in mouse ES cells, the role of FGF signaling and Sox2 levels on heterogeneous Nanog expression was examined. Interference with FGF or ERK activity by genetic ablation or signal inhibition, promoted high, homogenous Nanog expression and enhanced self-renewal. This is consistent with reports showing that similar manipulations reduced the ability of ES cells to commit to differentiation. Moreover, ES cells with reduced Sox2 levels displayed greater heterogeneity for Nanog expression than wild-type ES cells. Pluripotency is lost in the mouse embryo around E8.5, however, the precise timing and mechanism involved in this process has not yet been defined. Here it is shown that pluripotency is extinguished at the onset of somitogenesis, coincident with reduced expression and chromatin accessibility of Oct4 and Nanog regulatory regions. Prior to somitogenesis, the expression of both Nanog and Oct4 is regionalized. Interestingly, pluripotency tracks the in vivo level of Oct4, this correlation does not hold true for Nanog. However, Nanog expression reports on pluripotent cells. Indeed, ectopic Oct4 expression in somitogenesis-stage tissue provokes rapid reopening of Oct4 and Nanog chromatin, Nanog re-expression and resuscitation of moribund pluripotency. Competence to re-activate the pluripotency network upon enforced Oct4 expression is gradually lost with the progression of embryonic development. ES cells and EpiSCs are two distinct pluripotent populations as they show differences in their ability to undergo clonal propagation, re-colonize embryos, growth factor responsiveness, morphology and gene regulatory networks. It is possible to harness this differential growth factor responsiveness to convert ES cells into EpiSCs. Conversely, EpiSC can be reverted back to ES cell pluripotency through the overexpression of a small number of transcription factors. The inter-conversion of ES cells and EpiSCs has been documented, but detailed analyses of the changes that occur during such transitions had not been performed. The current work…
Subjects/Keywords: 571.6; pluripotency; ES cells
…38
2.1 Culture and manipulation of mouse ES cells… …39
2.1.1.4 Reagents for passaging of ES cells… …41
2.1.3 Long-term storage of mouse ES cells… …43
2.1.4 Thawing mouse ES cells… …43
2.1.5
Transfection of DNA into mouse ES cells…
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Manager
APA (6th Edition):
Osorno Hernandez, C. R. (2013). Transcription factor heterogeneity in epiblast pluripotency. (Doctoral Dissertation). University of Edinburgh. Retrieved from http://hdl.handle.net/1842/8081
Chicago Manual of Style (16th Edition):
Osorno Hernandez, Carlos Rodrigo. “Transcription factor heterogeneity in epiblast pluripotency.” 2013. Doctoral Dissertation, University of Edinburgh. Accessed September 17, 2019.
http://hdl.handle.net/1842/8081.
MLA Handbook (7th Edition):
Osorno Hernandez, Carlos Rodrigo. “Transcription factor heterogeneity in epiblast pluripotency.” 2013. Web. 17 Sep 2019.
Vancouver:
Osorno Hernandez CR. Transcription factor heterogeneity in epiblast pluripotency. [Internet] [Doctoral dissertation]. University of Edinburgh; 2013. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/1842/8081.
Council of Science Editors:
Osorno Hernandez CR. Transcription factor heterogeneity in epiblast pluripotency. [Doctoral Dissertation]. University of Edinburgh; 2013. Available from: http://hdl.handle.net/1842/8081
University of Edinburgh
12.
Zhang, Jingchao.
Functional analysis of the role of the Nanog tryptophan repeat in ES cells.
Degree: PhD, 2016, University of Edinburgh
URL: http://hdl.handle.net/1842/15972
► Nanog is a transcription factor that plays a central part in the gene regulatory network that maintains and induces pluripotency of embryonic stem cells (ESCs).…
(more)
▼ Nanog is a transcription factor that plays a central part in the gene regulatory network that maintains and induces pluripotency of embryonic stem cells (ESCs). However, the molecular basis by which Nanog achieves its functions is not fully understood. At the centre of C-terminal domain of Nanog a tryptophan repeat (WR) is located, comprising 10 penta-peptide repeats each starting with a tryptophan. A mutant form of Nanog (Nanog-W10A) in which all 10 tryptophan residues have been substituted by alanine has an impaired capacity to drive LIF-independent self-renewal and a reduced efficiency in reprogramming primed epiblast stem cells to naïve pluripotency. To understand how the WR contributes to Nanog function, Nanog-W10A-ERT2 was introduced into Nanog null cells. Upon hydroxytamoxifen addition, the Nanog-ERT2 fusion proteins were detected on chromatin within 1 hour, allowing a comparison of genome-wide transcriptional responses to Nanog and Nanog-W10A by microarray. When treated with LIF, Nanog-W10A can activate most of Nanog targets as efficiently as Nanog. In contrast, Nanog-W10A did not efficiently repress most Nanog targets, including Otx2 and Tcf15 that were previously suggested to prime ESCs for differentiation. The microarray experiments performed in the absence of LIF signalling showed that Nanog and LIF co-regulate an extensive list of targets, including Klf4 and Mras. When LIF is absent, wildtype Nanog can still activate pro-self-renewal factors, including Esrrb and repress differentiation-priming factor, such as Tcf15 and Otx2. In contrast, in the absence of LIF, the activation of pro-self-renewal factors Klf4 and Mras is reduced. In addition, activation of Esrrb by Nanog-W10A induction delays but does not prevent differentiation. These effects allow the de-repression of Otx2 and Tcf15 by Nanog-W10A to dominate. Therefore, the function of Nanog is not only mediated by the activation of pro-self-renewal genes, but also repression of pro-differentiation signals. The functional significance of the repression of Nanog targets was further exemplified by the robust capacity of Otx2 to dominate over the self-renewal signals and to drive differentiation. The Otx2 protein is a direct interacting partner of Nanog that binds the Nanog WR tryptophan residues. The previously identified Otx2 “tail domain” comprises two imperfectly aligned repeats and aromatic residues of each repeat align with aromatic residues of the Sox2 “SXS/TY” motif previously identified to mediate the interaction between Sox2 and Nanog. Aromatic residues of Otx2 were demonstrated to directly interact with both Nanog and Sox2. The interactions between Otx2, Nanog and Sox2 are essential for Otx2 functions in driving ESCs differentiation, as Otx2 mutants with alanine substitutions of the aromatic residues in both or either of the repeats have reduced efficiency to drive differentiation. As Nanog and Sox2 may co-occupy many loci important in maintaining ESC self-renewal, Otx2 may be able to “read” the Nanog/Sox2 co-binding sites to dissolve the…
Subjects/Keywords: 616.02; ES cells; Nanog; Otx2; tryptophan repeats; pluripotency
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CSE |
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APA (6th Edition):
Zhang, J. (2016). Functional analysis of the role of the Nanog tryptophan repeat in ES cells. (Doctoral Dissertation). University of Edinburgh. Retrieved from http://hdl.handle.net/1842/15972
Chicago Manual of Style (16th Edition):
Zhang, Jingchao. “Functional analysis of the role of the Nanog tryptophan repeat in ES cells.” 2016. Doctoral Dissertation, University of Edinburgh. Accessed September 17, 2019.
http://hdl.handle.net/1842/15972.
MLA Handbook (7th Edition):
Zhang, Jingchao. “Functional analysis of the role of the Nanog tryptophan repeat in ES cells.” 2016. Web. 17 Sep 2019.
Vancouver:
Zhang J. Functional analysis of the role of the Nanog tryptophan repeat in ES cells. [Internet] [Doctoral dissertation]. University of Edinburgh; 2016. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/1842/15972.
Council of Science Editors:
Zhang J. Functional analysis of the role of the Nanog tryptophan repeat in ES cells. [Doctoral Dissertation]. University of Edinburgh; 2016. Available from: http://hdl.handle.net/1842/15972
University of Rochester
13.
Gabay, Meital.
Ric-8 Proteins Are Molecular Chaperones Required For The
Biogenesis G Protein Alpha Subunits.
Degree: PhD, 2013, University of Rochester
URL: http://hdl.handle.net/1802/26791
► Ric-8A (resistance to inhibitors of cholinesterase 8A) and Ric-8B are guanine nucleotide exchange factors that enhance different heterotrimeric G protein signaling pathways by unknown mechanisms.…
(more)
▼ Ric-8A (resistance to inhibitors of cholinesterase
8A) and Ric-8B are guanine nucleotide
exchange factors that
enhance different heterotrimeric G protein signaling pathways by
unknown mechanisms. Because transgenic disruption of Ric-8A or
Ric-8B in mice caused
early embryonic lethality, we derived viable
Ric-8A– or Ric-8B–deleted embryonic stem
(ES) cell lines from
blastocysts of these mice.
We observed pleiotropic G protein
signaling defects in Ric-8A-/- ES cells, which
resulted from
reduced steady-state amounts of Gαi, Gαq, and Gα13 proteins to
<5% of
those of wild-type cells. Only the amount of Gαs was
reduced substantially in Ric-8B-/-
cells. The plasma membrane
residence of G proteins persisted in the absence of Ric-8 but
was
markedly reduced compared to that in wild-type cells. Endogenous
Gαi and Gαq
were efficiently translated in Ric-8A-/- cells but
integrated into endomembranes poorly;
however, the reduced amounts
of Gα subunits that reached the membrane still bound to
nascent
Gβγ. Interestingly, the Gαi and Gαq proteins that remained soluble
in the Ric-8A-
/- cells, had a higher apparent molecular weight,
suggestive towards the presence of a
putative post translational
modification (PTM) that is revealed when Ric-8A is absent. In
addition, Gαi, Gαq, and Gβ1 proteins exhibited accelerated rates of
degradation in Ric-
8A-/- cells compared to those in wild-type
cells.
Together, these data suggest that Ric-8 proteins are
molecular chaperones
required for the initial association of
nascent Gα subunits with cellular membranes. From
further
investigation of the slower migrating Gα proteins we concluded that
in the
absence of Ric-8 proteins, newly synthesized Gα proteins
are not properly folded and
become resistant to an artifactual
proteolytic event that occurs by cellular proteases. In
addition,
we found that Ric-8A is natively phosphorylation in mammalian
cells, however
some site directed mutations of predicted
phosphorylated sites resulted in full rescue of G
protein
expression while others generated a Ric-8A protein that was
unstable when
expressed in ES or insect cells. We believe that
phosphorylation or dephosphorylation of
Ric-8A has a functional
purpose that can reveal further mechanistic information in
regards
to the folding and initial trafficking of Gα proteins, as well as
the discovery of
cellular Ric-8 activators.
Subjects/Keywords: Ric-8; G Proteins; ES Cells; Folding; Chaperon
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APA (6th Edition):
Gabay, M. (2013). Ric-8 Proteins Are Molecular Chaperones Required For The
Biogenesis G Protein Alpha Subunits. (Doctoral Dissertation). University of Rochester. Retrieved from http://hdl.handle.net/1802/26791
Chicago Manual of Style (16th Edition):
Gabay, Meital. “Ric-8 Proteins Are Molecular Chaperones Required For The
Biogenesis G Protein Alpha Subunits.” 2013. Doctoral Dissertation, University of Rochester. Accessed September 17, 2019.
http://hdl.handle.net/1802/26791.
MLA Handbook (7th Edition):
Gabay, Meital. “Ric-8 Proteins Are Molecular Chaperones Required For The
Biogenesis G Protein Alpha Subunits.” 2013. Web. 17 Sep 2019.
Vancouver:
Gabay M. Ric-8 Proteins Are Molecular Chaperones Required For The
Biogenesis G Protein Alpha Subunits. [Internet] [Doctoral dissertation]. University of Rochester; 2013. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/1802/26791.
Council of Science Editors:
Gabay M. Ric-8 Proteins Are Molecular Chaperones Required For The
Biogenesis G Protein Alpha Subunits. [Doctoral Dissertation]. University of Rochester; 2013. Available from: http://hdl.handle.net/1802/26791
University of Bath
14.
Sanchez Ripoll, Yolanda.
Glycogen Synthase Kinase 3 (GSK-3) involvement in regulation of mouse embryonic stem cell fate.
Degree: PhD, 2011, University of Bath
URL: https://researchportal.bath.ac.uk/en/studentthesis/glycogen-synthase-kinase-3-gsk3-involvement-in-regulation-of-mouse-embryonic-stem-cell-fate(55f41abe-2856-455a-a76f-fd91e5034eea).html
;
https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.550616
Subjects/Keywords: 616.02774; ES cells; GSK-3
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APA (6th Edition):
Sanchez Ripoll, Y. (2011). Glycogen Synthase Kinase 3 (GSK-3) involvement in regulation of mouse embryonic stem cell fate. (Doctoral Dissertation). University of Bath. Retrieved from https://researchportal.bath.ac.uk/en/studentthesis/glycogen-synthase-kinase-3-gsk3-involvement-in-regulation-of-mouse-embryonic-stem-cell-fate(55f41abe-2856-455a-a76f-fd91e5034eea).html ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.550616
Chicago Manual of Style (16th Edition):
Sanchez Ripoll, Yolanda. “Glycogen Synthase Kinase 3 (GSK-3) involvement in regulation of mouse embryonic stem cell fate.” 2011. Doctoral Dissertation, University of Bath. Accessed September 17, 2019.
https://researchportal.bath.ac.uk/en/studentthesis/glycogen-synthase-kinase-3-gsk3-involvement-in-regulation-of-mouse-embryonic-stem-cell-fate(55f41abe-2856-455a-a76f-fd91e5034eea).html ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.550616.
MLA Handbook (7th Edition):
Sanchez Ripoll, Yolanda. “Glycogen Synthase Kinase 3 (GSK-3) involvement in regulation of mouse embryonic stem cell fate.” 2011. Web. 17 Sep 2019.
Vancouver:
Sanchez Ripoll Y. Glycogen Synthase Kinase 3 (GSK-3) involvement in regulation of mouse embryonic stem cell fate. [Internet] [Doctoral dissertation]. University of Bath; 2011. [cited 2019 Sep 17].
Available from: https://researchportal.bath.ac.uk/en/studentthesis/glycogen-synthase-kinase-3-gsk3-involvement-in-regulation-of-mouse-embryonic-stem-cell-fate(55f41abe-2856-455a-a76f-fd91e5034eea).html ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.550616.
Council of Science Editors:
Sanchez Ripoll Y. Glycogen Synthase Kinase 3 (GSK-3) involvement in regulation of mouse embryonic stem cell fate. [Doctoral Dissertation]. University of Bath; 2011. Available from: https://researchportal.bath.ac.uk/en/studentthesis/glycogen-synthase-kinase-3-gsk3-involvement-in-regulation-of-mouse-embryonic-stem-cell-fate(55f41abe-2856-455a-a76f-fd91e5034eea).html ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.550616
University of Cincinnati
15.
Fischer, Jared Michael.
Mouse Models of Mutation in vivo.
Degree: PhD, Medicine : Molecular Genetics, Biochemistry, and
Microbiology, 2008, University of Cincinnati
URL: http://rave.ohiolink.edu/etdc/view?acc_num=ucin1227214862
► Mutation is a driving factor in many diseases, in particular cancer, but also diseases such as atherosclerosis. Mutation has been studied in human blood…
(more)
▼ Mutation is a driving factor in many
diseases, in particular cancer, but also diseases such as
atherosclerosis. Mutation has been studied in human blood
cells and
other cell types that can be cultured. These mutation studies in
humans have revealed increased mutation based on chemical exposure
or tumor formation, but correlations between a particular gene or
chemical are difficult due to the varied nature of the human
population. Therefore mouse models of mutation have been developed
since mice can be inbred and a particular gene can be isolated and
studied. There are many different mouse models of
mutation, each with its own advantages and disadvantages. Some
mouse models of mutation can measure a wide range of mutations but
only in cell types that can be cultured. Other mouse models of
mutation can measure a smaller range of mutations in all tissues,
but the specific cell type is unknown. Still other mouse models of
mutation can measure only a specific type of mutation, but in many
tissues and in specific cell types. There is not yet a mouse that
can measure all mutations, in all tissues, and in specific cell
types. I have studied two different mouse models
of mutation: G11 PLAP and APRT. The G11 PLAP assay measures only
one type of mutation (a frameshift), but the specific cell types
that are mutated can be determined. The APRT assay measures many
different kinds of mutation, but only in cell types that can be
cultured. The G11 PLAP assay showed that not only
is there increased mutation upon chemical treatment and increasing
age, but there is also an increase in proliferation. A cell that is
mutated for G11 PLAP does not have a growth advantage; therefore
PLAP can be an independent marker for cell proliferation. It was
surprising to find an increase in proliferation with increasing
age, since most organs are thought to have a low mitotic index in
adulthood. The utility of the G11 PLAP assay to not only measure
mutation, but also proliferation, has led me to the design of
better mouse models that can detect of wider range of mutations,
but still be <i>in situ</i>. I have developed a
potential mouse model of mutation that can detect of wider range of
mutations, but still be <i>in
situ</i>. The shYFP mouse model has a siRNA
on one homologue and a fluorescent protein on the other homologue.
A normal cell is dim, but upon loss of the siRNA, the cell becomes
bright. This phenomenon was seen in both embryonic stem
cells and
intestinal epithelia. The loss of the siRNA was found to be from
large chromosomal events, but smaller mutation events could also
lead to loss of the siRNA. This is the first mouse model to measure
many different mutational events <i>in situ</i>. I have
continued to work on perfecting the model with slight variations
with the ultimate goal to make a mouse that can measure many
different kinds of mutation in any cell both rapidly and
easily.
Advisors/Committee Members: Stringer, James (Committee Chair).
Subjects/Keywords: Cellular Biology; Genetics; Molecular Biology; PLAP; cells; mice; Mutation; APRT; arsenic; ES cells
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APA ·
Chicago ·
MLA ·
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Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Fischer, J. M. (2008). Mouse Models of Mutation in vivo. (Doctoral Dissertation). University of Cincinnati. Retrieved from http://rave.ohiolink.edu/etdc/view?acc_num=ucin1227214862
Chicago Manual of Style (16th Edition):
Fischer, Jared Michael. “Mouse Models of Mutation in vivo.” 2008. Doctoral Dissertation, University of Cincinnati. Accessed September 17, 2019.
http://rave.ohiolink.edu/etdc/view?acc_num=ucin1227214862.
MLA Handbook (7th Edition):
Fischer, Jared Michael. “Mouse Models of Mutation in vivo.” 2008. Web. 17 Sep 2019.
Vancouver:
Fischer JM. Mouse Models of Mutation in vivo. [Internet] [Doctoral dissertation]. University of Cincinnati; 2008. [cited 2019 Sep 17].
Available from: http://rave.ohiolink.edu/etdc/view?acc_num=ucin1227214862.
Council of Science Editors:
Fischer JM. Mouse Models of Mutation in vivo. [Doctoral Dissertation]. University of Cincinnati; 2008. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=ucin1227214862
University of Toronto
16.
Norman, Andreea.
Characterization of ES Cell-derived Cortical Radial Precursor Differentiation.
Degree: 2010, University of Toronto
URL: http://hdl.handle.net/1807/25879
► Murine neural precursor cells have been a well studied model for neural cell fate determination and stem cell function both in vivo and in primary…
(more)
▼ Murine neural precursor cells have been a well studied model for neural cell fate determination and stem cell function both in vivo and in primary culture. However, factors such as cell number, the presence of multiple cell populations and of niche intrinsic factors made it difficult to dissect the mechanisms regulating cortical development. To overcome this issue, we have developed a culture system where mouse embryonic stem cells (ES) are differentiated to cortical radial precursors through retinoic acid treatment of embryoid bodies. One day after plating in neural differentiation conditions, ~70% of cells in the culture are cortical radial precursors (RPs) as indicated by the definitive cortical marker Emx1, and over 8 days in culture, these RPs differentiate to pyramidal glutamatergic neurons of the cortex mimicking in vivo development. Astrocyte differentiation can be observed later as the culture progresses, which again mimics the typical timed genesis of cells in the cortex. The stem cell properties and cell fate of these RPs can be manipulated with growth factors in culture as they are in vivo. In particular, FGF2 promotes proliferation and survival, while ciliary neurotrophic factor (CNTF) induces precocious astrocyte formation. Thus, our ES-derived cortical RP cultures can serve as an alternate and complementary in vitro model to examine neural precursor biology during early development.
MAST
Advisors/Committee Members: Miller, Freda, Molecular and Medical Genetics.
Subjects/Keywords: cortical precursors; neural stem cells; mouse ES cells; corticogenesis; in vitro differentiation; retinoic acid; 0317
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APA (6th Edition):
Norman, A. (2010). Characterization of ES Cell-derived Cortical Radial Precursor Differentiation. (Masters Thesis). University of Toronto. Retrieved from http://hdl.handle.net/1807/25879
Chicago Manual of Style (16th Edition):
Norman, Andreea. “Characterization of ES Cell-derived Cortical Radial Precursor Differentiation.” 2010. Masters Thesis, University of Toronto. Accessed September 17, 2019.
http://hdl.handle.net/1807/25879.
MLA Handbook (7th Edition):
Norman, Andreea. “Characterization of ES Cell-derived Cortical Radial Precursor Differentiation.” 2010. Web. 17 Sep 2019.
Vancouver:
Norman A. Characterization of ES Cell-derived Cortical Radial Precursor Differentiation. [Internet] [Masters thesis]. University of Toronto; 2010. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/1807/25879.
Council of Science Editors:
Norman A. Characterization of ES Cell-derived Cortical Radial Precursor Differentiation. [Masters Thesis]. University of Toronto; 2010. Available from: http://hdl.handle.net/1807/25879
Univerzitet u Beogradu
17.
Mitić, Ivana D., 1981-.
Modulacija eksperimentalnog autoimunskog
encefalomijelitisa primenom ekskretorno-sekretornih antigena
parazita Trichinella spiralis.
Degree: Medicinski fakultet, 2016, Univerzitet u Beogradu
URL: https://fedorabg.bg.ac.rs/fedora/get/o:14182/bdef:Content/get
► Medicina - Imunologija / Medicine - Immunology
Savremena istraživanja na eksperimentalnim modelima autoimunskih bolesti ukazuju da infekcija helmintom Trichinella spiralis (T. spiralis), inhibira nastanak ili…
(more)
▼ Medicina - Imunologija / Medicine -
Immunology
Savremena istraživanja na eksperimentalnim modelima
autoimunskih bolesti ukazuju da infekcija helmintom Trichinella
spiralis (T. spiralis), inhibira nastanak ili ublažava simptome
autoimunskih poremećaja kao što su eksperimentalni autoimunski
encefalomijelitis (EAE), eksperimentalni kolitis i dijabetes
melitus tip 1. Tokom infekcije parazitom T. spiralis
ekskretorno-sekretorni metabolički produkti mišićnih larvi (ES L1)
su ključni za interakciju sa organizmom domaćina. Stoga je
pretpostavka da su upravo ES L1 produkti odgovorni za modulaciju
toka autoimunskih oboljenja. Međutim, tačna uloga ES L1 antigena
ovog parazita u modulaciji bolesti, kao i mehanizmi te modulacije
nisu još uvek u potpunosti razjašnjeni. Predmet ovog istraživanja
bilo je ispitivanje uticaja direktne primene ES L1 produkata kao i
dendritskih ćelija (DC) stimulisanih ovim antigenima na tok i
modulaciju EAE-a kod DA pacova, sa ciljem da se utvrde mehanizmi
imunomodulacije kao i uloga dendritskih ćelija u njima. Ovde
prikazani rezultati, dobijeni u in vitro studijama na modelu DC
izolovanih iz kostne srži DA pacova, potvrdili su da ES antigeni T.
spiralis dovode do delimičnog sazrevanja DC koje se mogu označiti
kao tolerogene DC koje se odlikuju niskom produkcijom
pro-inflamatornog citokina IL-12p70 i povećanom produkcijom
anti-inflamatornog citokina IL-10. U ovom radu prvi put je
ustanovljeno da primena DC stimulisanih ES L1 antigenima T.
spiralis (DC/ES L1) ima inhibitorni efekat na razvoj EAE-a kod DA
pacova kao i da je ovaj efekat dozno zavisan. Najveći uticaj na
smanjenje težine bolesti ostvaren je primenom doze od 5 x 106 DC/ES
L1. Mehanizam koji je u osnovi ovog fenomena zasniva se na
polarizaciji imunskog odgovora u pravcu Th2 i regulatornog tipa
praćenog povećanom produkcijom Th2 i anti-inflamatornih citokina
(IL-4, IL-10 i TGF-β), a smanjenom produkcijom pro-inflamatornih
citokina IFN-γ i IL-17. Takođe, dolazi do povećanja procentualne
zastupljenosti CD4+CD25+Foxp3+ regulatornih T-limfocita (Treg) na
sistemskom nivou u slezini i na nivou ciljnog organa, u
CNS-u...
Advisors/Committee Members: Marković, Miloš, 1969-.
Subjects/Keywords: Trichinella spiralis; EAE; ES L1; immune response;
dendritic cells; regulatory T lymphocytes
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APA ·
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APA (6th Edition):
Mitić, Ivana D., 1. (2016). Modulacija eksperimentalnog autoimunskog
encefalomijelitisa primenom ekskretorno-sekretornih antigena
parazita Trichinella spiralis. (Thesis). Univerzitet u Beogradu. Retrieved from https://fedorabg.bg.ac.rs/fedora/get/o:14182/bdef:Content/get
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Mitić, Ivana D., 1981-. “Modulacija eksperimentalnog autoimunskog
encefalomijelitisa primenom ekskretorno-sekretornih antigena
parazita Trichinella spiralis.” 2016. Thesis, Univerzitet u Beogradu. Accessed September 17, 2019.
https://fedorabg.bg.ac.rs/fedora/get/o:14182/bdef:Content/get.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Mitić, Ivana D., 1981-. “Modulacija eksperimentalnog autoimunskog
encefalomijelitisa primenom ekskretorno-sekretornih antigena
parazita Trichinella spiralis.” 2016. Web. 17 Sep 2019.
Vancouver:
Mitić, Ivana D. 1. Modulacija eksperimentalnog autoimunskog
encefalomijelitisa primenom ekskretorno-sekretornih antigena
parazita Trichinella spiralis. [Internet] [Thesis]. Univerzitet u Beogradu; 2016. [cited 2019 Sep 17].
Available from: https://fedorabg.bg.ac.rs/fedora/get/o:14182/bdef:Content/get.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Mitić, Ivana D. 1. Modulacija eksperimentalnog autoimunskog
encefalomijelitisa primenom ekskretorno-sekretornih antigena
parazita Trichinella spiralis. [Thesis]. Univerzitet u Beogradu; 2016. Available from: https://fedorabg.bg.ac.rs/fedora/get/o:14182/bdef:Content/get
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
University of Lund
18.
Miskinyte, Giedre.
Generation of cortical neurons through reprogramming
technology.
Degree: 2018, University of Lund
URL: http://lup.lub.lu.se/record/d960f763-7d64-44c6-9574-c623790a429d
;
http://portal.research.lu.se/ws/files/54878264/Kvist_kappa_G5_hela_avh.pdf
► The human cortex is affected by several debilitating acute and chronic neurodegenerative disorders such as stroke, traumatic brain injury, amyotrophic lateral sclerosis and Alzheimer’s disease,…
(more)
▼ The human cortex is affected by several
debilitating acute and chronic neurodegenerative disorders such as
stroke, traumatic brain injury, amyotrophic lateral sclerosis and
Alzheimer’s disease, which target specific types of cortical
neurons. Emerging evidence indicates that stem cells and
reprogrammed cells can be used to generate human cortical neurons
both for cell replacement by transplantation, and for disease
modeling and drug screening. Several laboratories have established
in vitro protocols for the derivation of excitatory pyramidal
neurons, the principal type of neuron in the adult cortex, from
human pluripotent stem cells (hPSCs). Efficient production of
corticofugal projection neurons (CfuPNs) from ES cells has also
been reported. Alternatively, human fibroblasts have been directly
converted into induced neuronal (iN) cells. However, cortical
neurons have not been produced by this method. Also, it remains to
be assessed how closely the derived cortical neurons resemble their
in vivo counterparts, as well as, whether the generated cells are
capable of integrating into human neuronal circuits.The work
presented in this thesis demonstrates that cortical neuronal cells
can be produced from different types of starting cells: human
fibroblasts, human ES cells and human iPSC-derived NSPCs.
Regardless the starting cell context and derivation protocol, we
have produced in all studies neuronal cells pyramidal in shape,
expressing key cortical markers and functional in vitro. More
importantly, we showed that these cells are capable of forming
synaptic connections with adult human cortical neurons. It remains
to be assessed whether cells derived in this thesis are capable of
projecting to correct brain regions in vivo. Nonetheless, by
providing the first evidence that cortical neurons derived here
integrate in adult host neural networks also in a human-to-human
grafting situation, this thesis represents an early but important
step in the clinical translation of neuronal replacement to promote
recovery in the injured brain.
Subjects/Keywords: Cortex; Reprogramming; human ES cells; transcription factor programming; cortical projection neurons; Human adult cortical slices
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APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Miskinyte, G. (2018). Generation of cortical neurons through reprogramming
technology. (Doctoral Dissertation). University of Lund. Retrieved from http://lup.lub.lu.se/record/d960f763-7d64-44c6-9574-c623790a429d ; http://portal.research.lu.se/ws/files/54878264/Kvist_kappa_G5_hela_avh.pdf
Chicago Manual of Style (16th Edition):
Miskinyte, Giedre. “Generation of cortical neurons through reprogramming
technology.” 2018. Doctoral Dissertation, University of Lund. Accessed September 17, 2019.
http://lup.lub.lu.se/record/d960f763-7d64-44c6-9574-c623790a429d ; http://portal.research.lu.se/ws/files/54878264/Kvist_kappa_G5_hela_avh.pdf.
MLA Handbook (7th Edition):
Miskinyte, Giedre. “Generation of cortical neurons through reprogramming
technology.” 2018. Web. 17 Sep 2019.
Vancouver:
Miskinyte G. Generation of cortical neurons through reprogramming
technology. [Internet] [Doctoral dissertation]. University of Lund; 2018. [cited 2019 Sep 17].
Available from: http://lup.lub.lu.se/record/d960f763-7d64-44c6-9574-c623790a429d ; http://portal.research.lu.se/ws/files/54878264/Kvist_kappa_G5_hela_avh.pdf.
Council of Science Editors:
Miskinyte G. Generation of cortical neurons through reprogramming
technology. [Doctoral Dissertation]. University of Lund; 2018. Available from: http://lup.lub.lu.se/record/d960f763-7d64-44c6-9574-c623790a429d ; http://portal.research.lu.se/ws/files/54878264/Kvist_kappa_G5_hela_avh.pdf
19.
SLIM MZOUGHI.
CHARACTERIZATION OF THE PR-DOMAIN PROTEIN PRDM15: ROLE IN DEVELOPMENT AND CANCER.
Degree: 2015, National University of Singapore
URL: http://scholarbank.nus.edu.sg/handle/10635/121262
Subjects/Keywords: PRDM15; ES cells; development; Transcription; chromatin
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APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
MZOUGHI, S. (2015). CHARACTERIZATION OF THE PR-DOMAIN PROTEIN PRDM15: ROLE IN DEVELOPMENT AND CANCER. (Thesis). National University of Singapore. Retrieved from http://scholarbank.nus.edu.sg/handle/10635/121262
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
MZOUGHI, SLIM. “CHARACTERIZATION OF THE PR-DOMAIN PROTEIN PRDM15: ROLE IN DEVELOPMENT AND CANCER.” 2015. Thesis, National University of Singapore. Accessed September 17, 2019.
http://scholarbank.nus.edu.sg/handle/10635/121262.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
MZOUGHI, SLIM. “CHARACTERIZATION OF THE PR-DOMAIN PROTEIN PRDM15: ROLE IN DEVELOPMENT AND CANCER.” 2015. Web. 17 Sep 2019.
Vancouver:
MZOUGHI S. CHARACTERIZATION OF THE PR-DOMAIN PROTEIN PRDM15: ROLE IN DEVELOPMENT AND CANCER. [Internet] [Thesis]. National University of Singapore; 2015. [cited 2019 Sep 17].
Available from: http://scholarbank.nus.edu.sg/handle/10635/121262.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
MZOUGHI S. CHARACTERIZATION OF THE PR-DOMAIN PROTEIN PRDM15: ROLE IN DEVELOPMENT AND CANCER. [Thesis]. National University of Singapore; 2015. Available from: http://scholarbank.nus.edu.sg/handle/10635/121262
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
20.
VINOTH KUMAR S/O JAYASEELAN.
ROLE OF HUMAN EMBRYONIC STEM CELLS AND DERIVED PROGENIES IN GENOTOXICITY TESTING.
Degree: 2011, National University of Singapore
URL: http://scholarbank.nus.edu.sg/handle/10635/26141
Subjects/Keywords: ES cells; Toxicology testing; Genotoxicity Testing
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to Zotero / EndNote / Reference
Manager
APA (6th Edition):
JAYASEELAN, V. K. S. (2011). ROLE OF HUMAN EMBRYONIC STEM CELLS AND DERIVED PROGENIES IN GENOTOXICITY TESTING. (Thesis). National University of Singapore. Retrieved from http://scholarbank.nus.edu.sg/handle/10635/26141
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
JAYASEELAN, VINOTH KUMAR S/O. “ROLE OF HUMAN EMBRYONIC STEM CELLS AND DERIVED PROGENIES IN GENOTOXICITY TESTING.” 2011. Thesis, National University of Singapore. Accessed September 17, 2019.
http://scholarbank.nus.edu.sg/handle/10635/26141.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
JAYASEELAN, VINOTH KUMAR S/O. “ROLE OF HUMAN EMBRYONIC STEM CELLS AND DERIVED PROGENIES IN GENOTOXICITY TESTING.” 2011. Web. 17 Sep 2019.
Vancouver:
JAYASEELAN VKS. ROLE OF HUMAN EMBRYONIC STEM CELLS AND DERIVED PROGENIES IN GENOTOXICITY TESTING. [Internet] [Thesis]. National University of Singapore; 2011. [cited 2019 Sep 17].
Available from: http://scholarbank.nus.edu.sg/handle/10635/26141.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
JAYASEELAN VKS. ROLE OF HUMAN EMBRYONIC STEM CELLS AND DERIVED PROGENIES IN GENOTOXICITY TESTING. [Thesis]. National University of Singapore; 2011. Available from: http://scholarbank.nus.edu.sg/handle/10635/26141
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Indian Institute of Science
21.
Verma, Isha.
Insights into Differentiation of Mouse Pluripotent Stem Cells to Neural Lineage.
Degree: 2016, Indian Institute of Science
URL: http://etd.iisc.ernet.in/handle/2005/2885
;
http://etd.ncsi.iisc.ernet.in/abstracts/3747/G27748-Abs.pdf
► Pluripotent stem cells (PSCs: ESCs and iPSCs) provide an excellent model system for studying neural development and function. These cells also serve as a reliable…
(more)
▼ Pluripotent stem
cells (PSCs: ESCs and iPSCs) provide an excellent model system for studying neural development and function. These
cells also serve as a reliable source of cell replacement for the treatment of various neurodegenerative diseases and disorders. In view of these applications of PSCs, multiple protocols have been developed to direct their differentiation into neural lineage. However, many of these protocols are limiting in terms of
(a) low efficiency of generation of neural
cells after long-term culture, (b) requirement of exogenous factors to induce and enhance neural differentiation and (c) supplementation of PSC culture medium with serum. Therefore, in the present study, attempts were made to achieve enhanced efficiency of neural differentiation of PSCs in the absence of exogenous molecules by employing a defined culture medium containing knockout serum replacement (KSR). KSR-based culture system was tested with our in-house-derived EGFP-transgenic ‘GS-2’
ES-cell and ‘N9’ iPS-cell lines and the wild-type ‘D3’
ES-cell line. In KSR medium, PSC-derived EBs predominantly generated neural
cells from their post-attachment outgrowths and the complexity of neural networks increased as the culture progressed. Molecular phenotyping of PSC-derived neural
cells was performed based on the expression of neural markers both at the mRNA and protein levels. qPCR analysis revealed the expression of markers corresponding to multiple neural cell types, including glutamatergic, GABAergic, cholinergic, serotonergic and dopaminergic neurons, astrocytes and oligodendrocytes, at various time points during the culture. RNA expression studies were confirmed via immunocytochemical analysis of the expression of neural markers. On day 15 of culture, FACS quantitation revealed the efficient generation of NES+ neural progenitors (~16-18%), MAP2+ mature neurons (~12-26%) and GFAP+ astrocytes (~30-63%) from the three PSC lines. Functional assessment of the generated neurons was performed by electrophysiological analysis of passive (RMP) and active (threshold, amplitude, FWHM and outward and inward currents) membrane properties. In order to investigate the role of default pathway in neural differentiation of PSCs in KSR medium, various strategies were employed. GS-2
ES-
cells were cultured in the presence of different serum-free supplements; predominant differentiation into neural lineage was achieved in the B27-supplemented medium. The supplementation of KSR medium with BMP4 failed to show any effect of neural differentiation of GS-2
ES-
cells. Also, EBs were switched between KSR- and FBS-supplemented culture conditions on day 2 or day 5 of culture. These experiments indicated that KSR medium promoted the generation of neural cell fates at the expense of differentiation to non-neural lineages. Interestingly, differentiation of P19 EC-
cells in KSR medium also resulted in the predominant neural differentiation. These experiments collectively suggested the importance of default pathway in neural differentiation of PSCs in KSR medium.…
Advisors/Committee Members: Seshagiri, Polani B.
Subjects/Keywords: Mouse Pluripotent Stem Cells; Neural Lineage; Neural Differentiation of PSCs; Neural Differentiation; ES-cells; Neural Cell; Genetics
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APA (6th Edition):
Verma, I. (2016). Insights into Differentiation of Mouse Pluripotent Stem Cells to Neural Lineage. (Thesis). Indian Institute of Science. Retrieved from http://etd.iisc.ernet.in/handle/2005/2885 ; http://etd.ncsi.iisc.ernet.in/abstracts/3747/G27748-Abs.pdf
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Verma, Isha. “Insights into Differentiation of Mouse Pluripotent Stem Cells to Neural Lineage.” 2016. Thesis, Indian Institute of Science. Accessed September 17, 2019.
http://etd.iisc.ernet.in/handle/2005/2885 ; http://etd.ncsi.iisc.ernet.in/abstracts/3747/G27748-Abs.pdf.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Verma, Isha. “Insights into Differentiation of Mouse Pluripotent Stem Cells to Neural Lineage.” 2016. Web. 17 Sep 2019.
Vancouver:
Verma I. Insights into Differentiation of Mouse Pluripotent Stem Cells to Neural Lineage. [Internet] [Thesis]. Indian Institute of Science; 2016. [cited 2019 Sep 17].
Available from: http://etd.iisc.ernet.in/handle/2005/2885 ; http://etd.ncsi.iisc.ernet.in/abstracts/3747/G27748-Abs.pdf.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Verma I. Insights into Differentiation of Mouse Pluripotent Stem Cells to Neural Lineage. [Thesis]. Indian Institute of Science; 2016. Available from: http://etd.iisc.ernet.in/handle/2005/2885 ; http://etd.ncsi.iisc.ernet.in/abstracts/3747/G27748-Abs.pdf
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Universiteit Utrecht
22.
Bekhuis, L.M.
Evaluation of Midbrain Dopaminergic Neuron Specification in Rat ES Cell Derived Neurons.
Degree: 2013, Universiteit Utrecht
URL: http://dspace.library.uu.nl:8080/handle/1874/281018
► Parkinson's disease (PD) is a chronic, neurodegenerative disorder, which arises as a result of the progressive loss of midbrain dopaminergic (mDA) neurons in the substantia…
(more)
▼ Parkinson's disease (PD) is a chronic, neurodegenerative disorder, which arises as a result of the progressive loss of midbrain dopaminergic (mDA) neurons in the substantia nigra. The severity of the symptoms negatively influences the patients’ quality of life and comprises an economic burden to the government. Although many treatments have been developed over time, these therapies only reduce symptoms and often have side effects. Since PD results from the loss of a specific cell type, it lends itself to cell replacement therapy. A very promising source for cell replacement therapy are embryonic stem (
ES)
cells. The use of rat
ES cells in this perspective, offers some advantages compared to the utilization of mouse, human or primate
ES cells. Rat
ES cells have been reported to differentiate into neurons by means of adjusting the level of MEK/ERK inhibition. With regard to PD, it would be interesting to know if these neurons have differentiated toward a midbrain or even mDA neuronal fate. During this project, the RT-PCR conditions for markers for midbrain specification and mature mDA neurons were optimized and neuronal samples, previously generated by modulation of the level of MEK/ERK inhibition, were evaluated for midbrain marker expression by RT-PCR analysis. Indeed markers of mDA neuronal specification were identified in rat
ES cell derived neurons. This result provides useful knowledge for future experiments.
Advisors/Committee Members: Roelen, B.A.J., Burdon, T., Meek, S..
Subjects/Keywords: Diergeneeskunde; Parkinson's disease; midbrain dopaminergic neurons; embryonic stem cells; rat ES cells; MEK/ERK; RT-PCR; midbrain development
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APA ·
Chicago ·
MLA ·
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CSE |
Export
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APA (6th Edition):
Bekhuis, L. M. (2013). Evaluation of Midbrain Dopaminergic Neuron Specification in Rat ES Cell Derived Neurons. (Masters Thesis). Universiteit Utrecht. Retrieved from http://dspace.library.uu.nl:8080/handle/1874/281018
Chicago Manual of Style (16th Edition):
Bekhuis, L M. “Evaluation of Midbrain Dopaminergic Neuron Specification in Rat ES Cell Derived Neurons.” 2013. Masters Thesis, Universiteit Utrecht. Accessed September 17, 2019.
http://dspace.library.uu.nl:8080/handle/1874/281018.
MLA Handbook (7th Edition):
Bekhuis, L M. “Evaluation of Midbrain Dopaminergic Neuron Specification in Rat ES Cell Derived Neurons.” 2013. Web. 17 Sep 2019.
Vancouver:
Bekhuis LM. Evaluation of Midbrain Dopaminergic Neuron Specification in Rat ES Cell Derived Neurons. [Internet] [Masters thesis]. Universiteit Utrecht; 2013. [cited 2019 Sep 17].
Available from: http://dspace.library.uu.nl:8080/handle/1874/281018.
Council of Science Editors:
Bekhuis LM. Evaluation of Midbrain Dopaminergic Neuron Specification in Rat ES Cell Derived Neurons. [Masters Thesis]. Universiteit Utrecht; 2013. Available from: http://dspace.library.uu.nl:8080/handle/1874/281018
Indian Institute of Science
23.
Verma, Isha.
Insights into Differentiation of Mouse Pluripotent Stem Cells to Neural Lineage.
Degree: 2016, Indian Institute of Science
URL: http://hdl.handle.net/2005/2885
► Pluripotent stem cells (PSCs: ESCs and iPSCs) provide an excellent model system for studying neural development and function. These cells also serve as a reliable…
(more)
▼ Pluripotent stem
cells (PSCs: ESCs and iPSCs) provide an excellent model system for studying neural development and function. These
cells also serve as a reliable source of cell replacement for the treatment of various neurodegenerative diseases and disorders. In view of these applications of PSCs, multiple protocols have been developed to direct their differentiation into neural lineage. However, many of these protocols are limiting in terms of
(a) low efficiency of generation of neural
cells after long-term culture, (b) requirement of exogenous factors to induce and enhance neural differentiation and (c) supplementation of PSC culture medium with serum. Therefore, in the present study, attempts were made to achieve enhanced efficiency of neural differentiation of PSCs in the absence of exogenous molecules by employing a defined culture medium containing knockout serum replacement (KSR). KSR-based culture system was tested with our in-house-derived EGFP-transgenic ‘GS-2’
ES-cell and ‘N9’ iPS-cell lines and the wild-type ‘D3’
ES-cell line. In KSR medium, PSC-derived EBs predominantly generated neural
cells from their post-attachment outgrowths and the complexity of neural networks increased as the culture progressed. Molecular phenotyping of PSC-derived neural
cells was performed based on the expression of neural markers both at the mRNA and protein levels. qPCR analysis revealed the expression of markers corresponding to multiple neural cell types, including glutamatergic, GABAergic, cholinergic, serotonergic and dopaminergic neurons, astrocytes and oligodendrocytes, at various time points during the culture. RNA expression studies were confirmed via immunocytochemical analysis of the expression of neural markers. On day 15 of culture, FACS quantitation revealed the efficient generation of NES+ neural progenitors (~16-18%), MAP2+ mature neurons (~12-26%) and GFAP+ astrocytes (~30-63%) from the three PSC lines. Functional assessment of the generated neurons was performed by electrophysiological analysis of passive (RMP) and active (threshold, amplitude, FWHM and outward and inward currents) membrane properties. In order to investigate the role of default pathway in neural differentiation of PSCs in KSR medium, various strategies were employed. GS-2
ES-
cells were cultured in the presence of different serum-free supplements; predominant differentiation into neural lineage was achieved in the B27-supplemented medium. The supplementation of KSR medium with BMP4 failed to show any effect of neural differentiation of GS-2
ES-
cells. Also, EBs were switched between KSR- and FBS-supplemented culture conditions on day 2 or day 5 of culture. These experiments indicated that KSR medium promoted the generation of neural cell fates at the expense of differentiation to non-neural lineages. Interestingly, differentiation of P19 EC-
cells in KSR medium also resulted in the predominant neural differentiation. These experiments collectively suggested the importance of default pathway in neural differentiation of PSCs in KSR medium.…
Advisors/Committee Members: Seshagiri, Polani B.
Subjects/Keywords: Mouse Pluripotent Stem Cells; Neural Lineage; Neural Differentiation of PSCs; Neural Differentiation; ES-cells; Neural Cell; Genetics
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Record Details
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Verma, I. (2016). Insights into Differentiation of Mouse Pluripotent Stem Cells to Neural Lineage. (Thesis). Indian Institute of Science. Retrieved from http://hdl.handle.net/2005/2885
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Verma, Isha. “Insights into Differentiation of Mouse Pluripotent Stem Cells to Neural Lineage.” 2016. Thesis, Indian Institute of Science. Accessed September 17, 2019.
http://hdl.handle.net/2005/2885.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Verma, Isha. “Insights into Differentiation of Mouse Pluripotent Stem Cells to Neural Lineage.” 2016. Web. 17 Sep 2019.
Vancouver:
Verma I. Insights into Differentiation of Mouse Pluripotent Stem Cells to Neural Lineage. [Internet] [Thesis]. Indian Institute of Science; 2016. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/2005/2885.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Verma I. Insights into Differentiation of Mouse Pluripotent Stem Cells to Neural Lineage. [Thesis]. Indian Institute of Science; 2016. Available from: http://hdl.handle.net/2005/2885
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Indian Institute of Science
24.
Abbey, Deepti.
Insights Into Molecular Regulation Of Cardiomyocyte Differentiation Of Mouse Pluripotent Stem Cells.
Degree: 2012, Indian Institute of Science
URL: http://hdl.handle.net/2005/2455
► Pluripotent stem cells (PSCs) are specialized cells, which have remarkable ability to maintain in an undifferentiated state and are capable of undergoing differentiation to three…
(more)
▼ Pluripotent stem
cells (PSCs) are specialized
cells, which have remarkable ability to maintain in an undifferentiated state and are capable of undergoing differentiation to three germ-layer lineage cell types, under differentiation-enabling conditions. PSCs include embryonic stem (
ES)-
cells, embryonal carcinoma (EC)-
cells and embryonic germ (EG)-
cells.
ES-
cells are derived from the inner cell mass (ICM) of day 3.5 blastocysts (mouse). On the other hand, EC- and EG-
cells have different source of origin and exhibit some differences in terms of their differentiation abilities and culture requirements. These PSCs act as an ideal in-vitro model system to study early mammalian development and cell differentiation and, they could potentially be used for experimental cell-based therapy for a number of diseases. However, one of the problems encountered is the immune rejection of transplanted
cells. For this, immune-matched induced pluripotent stem (iPS)-
cells have been derived from somatic
cells, by forced expression of a few stemness genes. Although, human PSCs lines are being experimented, their cell-therapeutic potential is still far from being thoroughly tested due to lack of our understanding regarding lineage-specific differentiation, homing and structural-functional integration of differentiated cell types in the host environment. To understand these mechanisms, it is desirable to have fluorescently-marked PSCs and their differentiated cell-types, which could facilitate experimental cell transplantation studies.
In this regard, our laboratory has earlier generated enhanced green fluorescent protein (EGFP)-expressing FVB/N transgenic ‘green’ mouse: GU-3 line (Devgan et al., 2003). This transgenic mouse has been an excellent source of intrinsically green fluorescent cell types. Recently, we have derived a ‘GS-2’
ES-cell line from the GU-3 mouse line (Singh et al., 2012). Additionally, we envisaged the need for developing an iPS-cell line from the GU-3 mouse and then use them for studying cell differentiation. Thus, aims of the study described in the thesis are to: (1) develop an experimental system to derive EGFP-expressing fluorescently-marked iPS-cell line from a genetically non-permissive FVB/N mouse strain, characterize the established iPS-cell line and achieve differentiation of various cell types from EGFP-expressing iPS-cell line; (2) to study differentiation phenomenon, in particular to cardiac lineage, using select-cardiogenesis
modulators and (3) to assess the gene-expression profiles and signaling system associated with cardiomyocyte differentiation of PSCs.
This thesis is divided into four chapters with the 1st chapter being a review of literature followed by three data chapters. In the chapter I of the thesis, a comprehensive up-to¬date review of literature is provided pertaining to PSCs, their classification, derivation strategies especially for reprogramming of somatic
cells for iPSC generation, their differentiation potential and characterization, particularly to cardiac lineage. Various molecular…
Advisors/Committee Members: Seshagiri, Polani B.
Subjects/Keywords: Molecular Regulation; Cardiac Differentiation; Cell Differentiation; Pluripotent Stem Cells; Embryonic Stem Cells; Embryonal Carcinoma Cells; Cardiomyocytes; Cardiomyocyte Differentiation; Mouse Pluripotent Stem Cells; Pluripotent Stem Cells (PSCs); Stem Cell Research; Embryonic Germ Cells; EC-cells; ES-cells; Molecular Biology
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APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Abbey, D. (2012). Insights Into Molecular Regulation Of Cardiomyocyte Differentiation Of Mouse Pluripotent Stem Cells. (Thesis). Indian Institute of Science. Retrieved from http://hdl.handle.net/2005/2455
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Abbey, Deepti. “Insights Into Molecular Regulation Of Cardiomyocyte Differentiation Of Mouse Pluripotent Stem Cells.” 2012. Thesis, Indian Institute of Science. Accessed September 17, 2019.
http://hdl.handle.net/2005/2455.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Abbey, Deepti. “Insights Into Molecular Regulation Of Cardiomyocyte Differentiation Of Mouse Pluripotent Stem Cells.” 2012. Web. 17 Sep 2019.
Vancouver:
Abbey D. Insights Into Molecular Regulation Of Cardiomyocyte Differentiation Of Mouse Pluripotent Stem Cells. [Internet] [Thesis]. Indian Institute of Science; 2012. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/2005/2455.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Abbey D. Insights Into Molecular Regulation Of Cardiomyocyte Differentiation Of Mouse Pluripotent Stem Cells. [Thesis]. Indian Institute of Science; 2012. Available from: http://hdl.handle.net/2005/2455
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Indian Institute of Science
25.
Singh, Gurbind.
Studies On Embryonic Stem Cells From Enhanced Green Fluorescent Protein Transgenic Mice : Induction Of Cardiomyocyte Differentiation.
Degree: 2011, Indian Institute of Science
URL: http://hdl.handle.net/2005/2116
► Genesis of life begins with the fusion of female and male haploid gametes through a process of fertilization leading to the formation of a diploid…
(more)
▼ Genesis of life begins with the fusion of female and male haploid gametes through a process of fertilization leading to the formation of a diploid cell, the zygote. This undergoes successive cleavage divisions forming 2-, 4- and 8- cell embryos and their individual
cells (blastomeres) are totipotent. As development proceeds, there is a gradual restriction in their totipotency, resulting in the generation of two distinct cell lineages i.e., the differentiated trophectoderm (TE)
cells and the undifferentiated, inner cell mass (ICM) during blastocyst morphogenesis (Rossant and Tam 2009). During the course of development, the ICM
cells can give rise to all cell types of an organism and can also provide embryonic stem (
ES)-
cells when cultured in vitro (Evan and Kaufman 1981).
ES-
cells are pluripotent
cells, having the ability to self-renew indefinitely and differentiate into all the three primary germ layers (ectoderm, mesoderm and endoderm) derived-cell types.
ES-
cells are an excellent developmental model system to understand basic mechanisms of self-renewal, cell differentiation and function of various genes in vitro and in vivo (Capecchi 2001). Importantly, their cell derivatives could potentially be used for experimental cell-based therapy for a number of diseases. Although, human
ES-cell lines have been successfully derived and differentiated to various cell types (Thomson et al., 1998; Odorico et al., 2001), their cell-therapeutic potential is far from being tested, in view of the lack of our understanding of lineage-specific differentiation, homing and structural-functional integration of differentiated cell types in the host environment. To understand these mechanisms, it is desirable to have fluorescently-marked
ES-
cells and their differentiated cell-types, which could facilitate experimental cell transplantation studies.
In this regard, our laboratory has earlier generated enhanced green fluorescent protein (EGFP)-expressing FVB/N transgenic ‘green’ mouse, under the control of ubiquitous chicken -actin promoter (Devgan et al., 2003). This transgenic mouse has been an excellent source of intrinsically green fluorescent cell types. We have been attempting to derive
ES-cell line from this transgenic mouse. Because the derivation of
ES-cell line is genetic strain-dependent, with some strains being relatively permissible for
ES-cell derivation while others are quite resistant (non permissive), it has been extremely difficult to derive
ES-cell line from the FVB/N mouse strain. There is a need to evolve experimental strategies to derive
ES-cell line from FVB/N mouse, a strain extensively used for transgenesis. Thus, the aims of the study described in the thesis are to: (1) develop an experimental system to derive EGFP-expressing fluorescently-marked
ES-cell line from a non-permissive FVB/N mouse strain; (2) characterize the established
ES-cell line; (3) achieve differentiation of various cell types from EGFP-expressing
ES-cell line and (4) understand role of FGF signaling in cardiac differentiation from the…
Advisors/Committee Members: Seshagiri, Polani B.
Subjects/Keywords: Experimental Research; Stem Cells; Transgenic Mice; Cardiomyocyte Differentiation; Embryonic Stem Cells; Enhanced Green Fluorescent Protein (EGFP); Fibroblast Growth Factor (FGF); Embryonic Stem Cell Line; ES-cells; Molecular Biology
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APA ·
Chicago ·
MLA ·
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CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Singh, G. (2011). Studies On Embryonic Stem Cells From Enhanced Green Fluorescent Protein Transgenic Mice : Induction Of Cardiomyocyte Differentiation. (Thesis). Indian Institute of Science. Retrieved from http://hdl.handle.net/2005/2116
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Singh, Gurbind. “Studies On Embryonic Stem Cells From Enhanced Green Fluorescent Protein Transgenic Mice : Induction Of Cardiomyocyte Differentiation.” 2011. Thesis, Indian Institute of Science. Accessed September 17, 2019.
http://hdl.handle.net/2005/2116.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Singh, Gurbind. “Studies On Embryonic Stem Cells From Enhanced Green Fluorescent Protein Transgenic Mice : Induction Of Cardiomyocyte Differentiation.” 2011. Web. 17 Sep 2019.
Vancouver:
Singh G. Studies On Embryonic Stem Cells From Enhanced Green Fluorescent Protein Transgenic Mice : Induction Of Cardiomyocyte Differentiation. [Internet] [Thesis]. Indian Institute of Science; 2011. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/2005/2116.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Singh G. Studies On Embryonic Stem Cells From Enhanced Green Fluorescent Protein Transgenic Mice : Induction Of Cardiomyocyte Differentiation. [Thesis]. Indian Institute of Science; 2011. Available from: http://hdl.handle.net/2005/2116
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
University of Southern California
26.
Andrianakos, Rosemary G.
Roles of Klf4 in embryonic stem cells.
Degree: MS, Biochemistry & Molecular Biology, 2008, University of Southern California
URL: http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll127/id/70866/rec/5646
► Human and mouse ES cells are defined by their ability to confer self-renewal and differentiate into various cell types, making them an invaluable tool for…
(more)
▼ Human and mouse
ES cells are defined by their ability
to confer self-renewal and differentiate into various cell types,
making them an invaluable tool for investigation of Stem cell
maintenance, differentiation, and reprogramming. Human
ES cells
have infinite potential for therapeutic applications.
Unfortunately, acquiring
ES cells is problematic, and reprogramming
somatic
cells to Induced Pluripotent Stem
Cells occurs by an
unknown mechanism.; This thesis establishes Klf4' s role in
ES cell
maintenance. Klf4 interacts with core
ES cell maintenance
transcription factors, Oct-4 and Sox-2, which bind to Klf4 's
C-terminal domain zinc fingers, and this binding is essential for
the collective function of the
ES-Cell maintenance network.
Moreover, this study contributes to the knowledge of dose dependant
cross-regulation of various factors in the stem cell maintenance
network, and provides a model for understanding how these factors
may confer
ES cell stability, self renewal, and
reprogramming.
Advisors/Committee Members: Lu, Wange (Committee Chair), Tokes, Zoltan A. (Committee Member), Stellwagen, Robert H. (Committee Member).
Subjects/Keywords: stem cells; reprogramming somatic cells; Klf4; Oct-4; Sox-2; differentiation of Es cells; pluripotency; ES cell maintenance and self renewal
Record Details
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APA ·
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MLA ·
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Export
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APA (6th Edition):
Andrianakos, R. G. (2008). Roles of Klf4 in embryonic stem cells. (Masters Thesis). University of Southern California. Retrieved from http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll127/id/70866/rec/5646
Chicago Manual of Style (16th Edition):
Andrianakos, Rosemary G. “Roles of Klf4 in embryonic stem cells.” 2008. Masters Thesis, University of Southern California. Accessed September 17, 2019.
http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll127/id/70866/rec/5646.
MLA Handbook (7th Edition):
Andrianakos, Rosemary G. “Roles of Klf4 in embryonic stem cells.” 2008. Web. 17 Sep 2019.
Vancouver:
Andrianakos RG. Roles of Klf4 in embryonic stem cells. [Internet] [Masters thesis]. University of Southern California; 2008. [cited 2019 Sep 17].
Available from: http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll127/id/70866/rec/5646.
Council of Science Editors:
Andrianakos RG. Roles of Klf4 in embryonic stem cells. [Masters Thesis]. University of Southern California; 2008. Available from: http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll127/id/70866/rec/5646
Dublin City University
27.
McKiernan, Eadaoin.
An investigation of differentiation programmes governing pancreatic cell development in vitro.
Degree: School of Biotechnology, 2006, Dublin City University
URL: http://doras.dcu.ie/18070/
► The murine embryonic stem (ES) cell line, ES-D3 was used in this project as a model for (a) investigating controversial aspects of current protocols used…
(more)
▼ The murine embryonic stem (
ES) cell line,
ES-D3 was used in this project as a model for (a) investigating controversial aspects of current protocols used for the derivation of (3 cell types from
ES cells and (b) investigating the potential to regulate the differentiation of
ES cells into endodermal derivatives 1 e 0-1 ike
cells, using extra-cellular factors previously associated with various aspects of pancreatic development.
Several groups have reported the derivation of insulin-expressing cell types from
ES cells. Many believe that these insulin-positive
cells were a result of insulin uptake from supplements used in the culture medium Others have suggested that the
ES-denved insulin-expressing
cells may be of a neuronal or extraembryonic lineage, rather than an endocrine lineage Using an adapted version of the Lumelsky protocol (Lumelsky et al, 2001), we demonstrated the induction of 3-dimensional clusters similar to those described in the original study, which expressed a range of pancreatic, extra-embryonic and neuronal transcripts. Insulin protein was detected in the differentiated clusters by ELISA. A novel method developed to detect insulin biosynthesis by the
cells, and analysis of Cpeptide expression, indicated that this insulin is probably derived from the culture medium, not from the differentiated
ES cell.
Culture of
ES cells in suspension leads to the formation of embryoid bodies (EBs). Treatment of these aggregates with retinoic acid (RA) was previously shown to induce endodermal differentiation We exposed RA_EBs to secondary treatment with agents associated with pancreatic endocrine differentiation, e g Activin A (AA), Sodium Butyrate (SB) and Betacellulin (BTC), with the aim of developing a novel protocol for the regulated derivation of insulin-expressing cell types. The resulting differentiated populations were analysed for transcript expression of a range of multi-lineage markers i e neuronal, myogenic, exocrine and endocrine pancreas, extra-embryonic and apoptotic markers. Immunohistochemistry methods were established and applied for the analysis of a selection of the chosen markers, following formal in-fixing and paraffinembedding. Applying 2-stage regulated differentiation protocols developedduring the course of the study, we demonstrated the derivation of an intermediate multi-potential population (RAJEBs) from undifferentiated
ES cells that preferentially gave rise to pancreatic endocrine insulin-expressing cell types in the presence of SB and neuronal-like cell types in the presence of AA or BTC.
Low passages of the MIN-6 insulinoma cell line are glucose-responsive whereas at high passage they become glucose non-responsive. This loss in the glucose stimulated insulin secretion (GSIS) phenotype correlates with changes in morphology, increased proliferation and increased alkaline phosphatase (AP) activity, suggesting that the loss of GSIS may be due to de-differentiation of the high passage MIN6
cells. To investigate this hypothesis further, gene expression differences between low passage and…
Advisors/Committee Members: Clynes, Martin, O'Driscoll, Lorraine.
Subjects/Keywords: Biotechnology; embryonic stem (ES) cells; pancreatic development; endodermal differentiation
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APA ·
Chicago ·
MLA ·
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CSE |
Export
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Manager
APA (6th Edition):
McKiernan, E. (2006). An investigation of differentiation programmes governing pancreatic cell development in vitro. (Thesis). Dublin City University. Retrieved from http://doras.dcu.ie/18070/
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
McKiernan, Eadaoin. “An investigation of differentiation programmes governing pancreatic cell development in vitro.” 2006. Thesis, Dublin City University. Accessed September 17, 2019.
http://doras.dcu.ie/18070/.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
McKiernan, Eadaoin. “An investigation of differentiation programmes governing pancreatic cell development in vitro.” 2006. Web. 17 Sep 2019.
Vancouver:
McKiernan E. An investigation of differentiation programmes governing pancreatic cell development in vitro. [Internet] [Thesis]. Dublin City University; 2006. [cited 2019 Sep 17].
Available from: http://doras.dcu.ie/18070/.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
McKiernan E. An investigation of differentiation programmes governing pancreatic cell development in vitro. [Thesis]. Dublin City University; 2006. Available from: http://doras.dcu.ie/18070/
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Kyoto University / 京都大学
28.
Shiraishi, Atsushi.
Generation of thalamic neurons from mouse embryonic stem cells : マウス胚性幹細胞からの視床神経の分化誘導.
Degree: 博士(医学), 2018, Kyoto University / 京都大学
URL: http://hdl.handle.net/2433/230993
;
http://dx.doi.org/10.14989/doctor.k20790
新制・課程博士
甲第20790号
医博第4290号
Subjects/Keywords: mouse ES cells; thalamus; caudal forebrain; self-organization; SFEBq
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APA (6th Edition):
Shiraishi, A. (2018). Generation of thalamic neurons from mouse embryonic stem cells : マウス胚性幹細胞からの視床神経の分化誘導. (Thesis). Kyoto University / 京都大学. Retrieved from http://hdl.handle.net/2433/230993 ; http://dx.doi.org/10.14989/doctor.k20790
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Shiraishi, Atsushi. “Generation of thalamic neurons from mouse embryonic stem cells : マウス胚性幹細胞からの視床神経の分化誘導.” 2018. Thesis, Kyoto University / 京都大学. Accessed September 17, 2019.
http://hdl.handle.net/2433/230993 ; http://dx.doi.org/10.14989/doctor.k20790.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Shiraishi, Atsushi. “Generation of thalamic neurons from mouse embryonic stem cells : マウス胚性幹細胞からの視床神経の分化誘導.” 2018. Web. 17 Sep 2019.
Vancouver:
Shiraishi A. Generation of thalamic neurons from mouse embryonic stem cells : マウス胚性幹細胞からの視床神経の分化誘導. [Internet] [Thesis]. Kyoto University / 京都大学; 2018. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/2433/230993 ; http://dx.doi.org/10.14989/doctor.k20790.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Shiraishi A. Generation of thalamic neurons from mouse embryonic stem cells : マウス胚性幹細胞からの視床神経の分化誘導. [Thesis]. Kyoto University / 京都大学; 2018. Available from: http://hdl.handle.net/2433/230993 ; http://dx.doi.org/10.14989/doctor.k20790
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Kyoto University
29.
Shiraishi, Atsushi.
Generation of thalamic neurons from mouse embryonic stem cells
.
Degree: 2018, Kyoto University
URL: http://hdl.handle.net/2433/230993
Subjects/Keywords: mouse ES cells;
thalamus;
caudal forebrain;
self-organization;
SFEBq
Record Details
Similar Records
Cite
Share »
Record Details
Similar Records
Cite
« Share
❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Shiraishi, A. (2018). Generation of thalamic neurons from mouse embryonic stem cells
. (Thesis). Kyoto University. Retrieved from http://hdl.handle.net/2433/230993
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Shiraishi, Atsushi. “Generation of thalamic neurons from mouse embryonic stem cells
.” 2018. Thesis, Kyoto University. Accessed September 17, 2019.
http://hdl.handle.net/2433/230993.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Shiraishi, Atsushi. “Generation of thalamic neurons from mouse embryonic stem cells
.” 2018. Web. 17 Sep 2019.
Vancouver:
Shiraishi A. Generation of thalamic neurons from mouse embryonic stem cells
. [Internet] [Thesis]. Kyoto University; 2018. [cited 2019 Sep 17].
Available from: http://hdl.handle.net/2433/230993.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Shiraishi A. Generation of thalamic neurons from mouse embryonic stem cells
. [Thesis]. Kyoto University; 2018. Available from: http://hdl.handle.net/2433/230993
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
30.
LAI ZHENYANG.
Transcriptome Study of Human Embryonic Stem Cells and Knockdown Study of a Pluripotency Marker, LIN28.
Degree: 2010, National University of Singapore
URL: http://scholarbank.nus.edu.sg/handle/10635/20462
Subjects/Keywords: human ES cells; pluripotency; LIN28; RNAi; post-transcriptional regulation
Record Details
Similar Records
Cite
Share »
Record Details
Similar Records
Cite
« Share
❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
ZHENYANG, L. (2010). Transcriptome Study of Human Embryonic Stem Cells and Knockdown Study of a Pluripotency Marker, LIN28. (Thesis). National University of Singapore. Retrieved from http://scholarbank.nus.edu.sg/handle/10635/20462
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
ZHENYANG, LAI. “Transcriptome Study of Human Embryonic Stem Cells and Knockdown Study of a Pluripotency Marker, LIN28.” 2010. Thesis, National University of Singapore. Accessed September 17, 2019.
http://scholarbank.nus.edu.sg/handle/10635/20462.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
ZHENYANG, LAI. “Transcriptome Study of Human Embryonic Stem Cells and Knockdown Study of a Pluripotency Marker, LIN28.” 2010. Web. 17 Sep 2019.
Vancouver:
ZHENYANG L. Transcriptome Study of Human Embryonic Stem Cells and Knockdown Study of a Pluripotency Marker, LIN28. [Internet] [Thesis]. National University of Singapore; 2010. [cited 2019 Sep 17].
Available from: http://scholarbank.nus.edu.sg/handle/10635/20462.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
ZHENYANG L. Transcriptome Study of Human Embryonic Stem Cells and Knockdown Study of a Pluripotency Marker, LIN28. [Thesis]. National University of Singapore; 2010. Available from: http://scholarbank.nus.edu.sg/handle/10635/20462
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
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Open Access Theses and Dissertations
