subject:(mammary organoids)

1. Hume, Robert David. Recapitulating mammary gland development and breast cancer cell migration in vitro using 3D engineered scaffolds.

Degree: PhD, 2018, University of Cambridge

URL: https://www.repository.cam.ac.uk/handle/1810/271099

► The adult mammary gland is comprised of a bi-layered epithelium of luminal and myoepithelial cells surrounded by an adipocyte-rich fat pad, a highly collagenous extra-cellular… (more)

▼ The adult mammary gland is comprised of a bi-layered epithelium of luminal and myoepithelial cells surrounded by an adipocyte-rich fat pad, a highly collagenous extra-cellular matrix (ECM) and a number of other stromal and endothelial cell types. Mammary stem cells (MaSCs) reside within the epithelium and these are capable of repopulating a mammary fat pad that is devoid of epithelium, upon transplantation. It was sought to recapitulate this process of MaSCs repopulating a fat pad using a synthetic fat pad, engineered from a collagen scaffold invested with adipocytes, to provide an in vitro 3D model. Fluorescently tagged murine Axin2-expressing cells were obtained from transgenic mice and seeded into these scaffolds and cultured, mimicking the process of fat pad repopulation. Immunohistochemical analysis demonstrated that Axin2+ myoepithelial cells were rarely capable of forming bi-layered structures that expressed correct myoepithelial localisation and resemblance to a luminal morphology. Breast tumours surrounded by anisotropic (directional) collagen fibres running perpendicular to the tumour boundary are more aggressive and associated with poor patient prognosis. To recapitulate this anisotropic collagen phenotype in vitro, an ice-templating technique was used to modify the structure of the collagen scaffolds producing both non-directional (isotropic) and anisotropic internal architectures. Tumour cells from various breast cancer cell lines were seeded into both isotropic and anisotropic scaffolds to investigate whether this approach could distinguish cell type-specific migratory ability and whether anisotropy affected migration efficiency. Following analysis by confocal microscopy and ImageJ, anisotropic scaffolds were observed to enhance the migratory potential of MDA-MB-231 breast cancer cells. These results highlight the importance of collagen alignment and provide a reproducible method to quantitatively measure cell migration in 3D for cells derived from different breast cancer subtypes. Building on these data, the protocol was adapted to permit the direct investigation of tumour biopsy material. Given the heterogeneity of breast tumours, it was considered important to maintain tumour architecture and stromal components. Thus, murine mammary tumour fragments from two different established mammary cancer models were utilised and cultured in anisotropic collagen scaffolds in the presence or absence of adipocytes to allow an investigation of their influence on tumour cell migration. Further experiments included addition of various therapeutic drugs followed by immunofluorescence microscopy coupled with an optical clearing technique. These data demonstrated the utility of the model in determining both the rate and capacity of tumour cells to migrate through the engineered stroma while shedding light also on the mode of migration. Moreover, the response of different mammary tumour types to chemotherapeutic drugs could be could be readily quantified. To humanize the fat pad for subsequent human tissue analysis,…

Subjects/Keywords: Breast cancer; Tissue Engineering; Mammary gland; Anisotropic; Anisotropy; Collagen; Adipocytes; 3D culture; Organoids

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Hume, R. D. (2018). Recapitulating mammary gland development and breast cancer cell migration in vitro using 3D engineered scaffolds. (Doctoral Dissertation). University of Cambridge. Retrieved from https://www.repository.cam.ac.uk/handle/1810/271099

Chicago Manual of Style (16^th Edition):

Hume, Robert David. “Recapitulating mammary gland development and breast cancer cell migration in vitro using 3D engineered scaffolds.” 2018. Doctoral Dissertation, University of Cambridge. Accessed March 03, 2021. https://www.repository.cam.ac.uk/handle/1810/271099.

MLA Handbook (7^th Edition):

Hume, Robert David. “Recapitulating mammary gland development and breast cancer cell migration in vitro using 3D engineered scaffolds.” 2018. Web. 03 Mar 2021.

Vancouver:

Hume RD. Recapitulating mammary gland development and breast cancer cell migration in vitro using 3D engineered scaffolds. [Internet] [Doctoral dissertation]. University of Cambridge; 2018. [cited 2021 Mar 03]. Available from: https://www.repository.cam.ac.uk/handle/1810/271099.

Council of Science Editors:

Hume RD. Recapitulating mammary gland development and breast cancer cell migration in vitro using 3D engineered scaffolds. [Doctoral Dissertation]. University of Cambridge; 2018. Available from: https://www.repository.cam.ac.uk/handle/1810/271099

University of Cambridge

2. Hume, Robert David. Recapitulating mammary gland development and breast cancer cell migration in vitro using 3D engineered scaffolds.

Degree: PhD, 2018, University of Cambridge

URL: https://doi.org/10.17863/CAM.18071 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.744466

► The adult mammary gland is comprised of a bi-layered epithelium of luminal and myoepithelial cells surrounded by an adipocyte-rich fat pad, a highly collagenous extra-cellular… (more)

▼ The adult mammary gland is comprised of a bi-layered epithelium of luminal and myoepithelial cells surrounded by an adipocyte-rich fat pad, a highly collagenous extra-cellular matrix (ECM) and a number of other stromal and endothelial cell types. Mammary stem cells (MaSCs) reside within the epithelium and these are capable of repopulating a mammary fat pad that is devoid of epithelium, upon transplantation. It was sought to recapitulate this process of MaSCs repopulating a fat pad using a synthetic fat pad, engineered from a collagen scaffold invested with adipocytes, to provide an in vitro 3D model. Fluorescently tagged murine Axin2-expressing cells were obtained from transgenic mice and seeded into these scaffolds and cultured, mimicking the process of fat pad repopulation. Immunohistochemical analysis demonstrated that Axin2+ myoepithelial cells were rarely capable of forming bi-layered structures that expressed correct myoepithelial localisation and resemblance to a luminal morphology. Breast tumours surrounded by anisotropic (directional) collagen fibres running perpendicular to the tumour boundary are more aggressive and associated with poor patient prognosis. To recapitulate this anisotropic collagen phenotype in vitro, an ice-templating technique was used to modify the structure of the collagen scaffolds producing both non-directional (isotropic) and anisotropic internal architectures. Tumour cells from various breast cancer cell lines were seeded into both isotropic and anisotropic scaffolds to investigate whether this approach could distinguish cell type-specific migratory ability and whether anisotropy affected migration efficiency. Following analysis by confocal microscopy and ImageJ, anisotropic scaffolds were observed to enhance the migratory potential of MDA-MB-231 breast cancer cells. These results highlight the importance of collagen alignment and provide a reproducible method to quantitatively measure cell migration in 3D for cells derived from different breast cancer subtypes. Building on these data, the protocol was adapted to permit the direct investigation of tumour biopsy material. Given the heterogeneity of breast tumours, it was considered important to maintain tumour architecture and stromal components. Thus, murine mammary tumour fragments from two different established mammary cancer models were utilised and cultured in anisotropic collagen scaffolds in the presence or absence of adipocytes to allow an investigation of their influence on tumour cell migration. Further experiments included addition of various therapeutic drugs followed by immunofluorescence microscopy coupled with an optical clearing technique. These data demonstrated the utility of the model in determining both the rate and capacity of tumour cells to migrate through the engineered stroma while shedding light also on the mode of migration. Moreover, the response of different mammary tumour types to chemotherapeutic drugs could be could be readily quantified. To humanize the fat pad for subsequent human tissue analysis,…

Subjects/Keywords: 616.99; Breast cancer; Tissue Engineering; Mammary gland; Anisotropic; Anisotropy; Collagen; Adipocytes; 3D culture; Organoids

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Hume, R. D. (2018). Recapitulating mammary gland development and breast cancer cell migration in vitro using 3D engineered scaffolds. (Doctoral Dissertation). University of Cambridge. Retrieved from https://doi.org/10.17863/CAM.18071 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.744466

Chicago Manual of Style (16^th Edition):

Hume, Robert David. “Recapitulating mammary gland development and breast cancer cell migration in vitro using 3D engineered scaffolds.” 2018. Doctoral Dissertation, University of Cambridge. Accessed March 03, 2021. https://doi.org/10.17863/CAM.18071 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.744466.

MLA Handbook (7^th Edition):

Hume, Robert David. “Recapitulating mammary gland development and breast cancer cell migration in vitro using 3D engineered scaffolds.” 2018. Web. 03 Mar 2021.

Vancouver:

Hume RD. Recapitulating mammary gland development and breast cancer cell migration in vitro using 3D engineered scaffolds. [Internet] [Doctoral dissertation]. University of Cambridge; 2018. [cited 2021 Mar 03]. Available from: https://doi.org/10.17863/CAM.18071 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.744466.

Council of Science Editors:

Hume RD. Recapitulating mammary gland development and breast cancer cell migration in vitro using 3D engineered scaffolds. [Doctoral Dissertation]. University of Cambridge; 2018. Available from: https://doi.org/10.17863/CAM.18071 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.744466

University of Cambridge

3. Harris, Olivia. Elucidation of mammary gland development: combination of novel techniques to study proliferation and stem cells.

Degree: PhD, 2019, University of Cambridge

URL: https://www.repository.cam.ac.uk/handle/1810/291054

► The mammary gland is a dynamic organ that undergoes many cycles of proliferation and death throughout both oestrus cycling and the gestation/lactation/involution cycle. Moreover, it… (more)

▼ The mammary gland is a dynamic organ that undergoes many cycles of proliferation and death throughout both oestrus cycling and the gestation/lactation/involution cycle. Moreover, it is a unique tissue in that the majority of its development occurs postnatally coincident with the production of ovarian hormones in puberty. This capacity of the mammary gland for rapid growth and regeneration has been attributed to mammary stem cells (MaSCs). However, despite extensive efforts over the past 60 years, definitive characterisation of the stem and progenitor cells of the mammary gland has yet to be achieved. A number of recent conflicting, lineage tracing studies have served only to fuel the fires of controversy, with previous characterisation of the mammary gland largely carried out using two-dimensional tissue sections. However, in order to fully appreciate the capacity of MaSCs and to maintain spatial information of the complex topological structure, the mammary gland must be investigated in its intact form. Accordingly, there is still much disagreement regarding the potency, capacity and location of MaSCs. Consequently, in order to unequivocally elucidate the MaSC hierarchy, a variety of novel techniques have been combined in this thesis. The first involves development and optimisation of optical tissue clearing techniques to allow the visualisation of the native mammary gland, in situ, and in three dimensions. To do so, a number of different optical tissue clearing methods have been assessed and combined with a variety of microscopy techniques to allow the multiple focal planes of the ductal network to be examined. These imaging techniques were then combined with two neutral lineage tracing models; the first, the Rosa26[CA]30 model, utilises stochastic continuous clonal labelling to allow for the fate tracking of the progeny from single functional stem and progenitor cells. The second unbiased lineage tracing approach, the Rosa26-Confetti model, allows for the mammary stem and progenitor progeny to be traced with precise timing, with the additional benefit of a multicolour reporter. Next, proliferation was examined in wholemount tissues to investigate the functional requirements for MaSCs, and their potential locations. Finally, these techniques have been combined with an ex vivo 3D organoid culture system to investigate the use of culture methods in examining mammary epithelial cell dynamics. By combination of these techniques, clonally marked regions can be investigated throughout the development of the mammary gland, from the formation of the embryonic mammary iii rudiment, to expansion of the ductal tree in puberty, and ultimately their fate in lactation and involution, where the mammary gland fulfils its evolutionary purpose. The mammary gland provides a unique opportunity to investigate epithelial development extra-embryonically that is not available in other tissues. Moreover, study of maintenance and turnover of this organ has important implications for other epithelial systems. Finally, elucidation of the…

Subjects/Keywords: stem cells; mammary gland; adult stem cells; lineage tracing; proliferation; breast; breast cancer; MaSC; MaSCs; mammary gland stem cells; mammary stem cells; confetti; CA30; organoids; 3D models; mammary gland organoids; epithelial biology; epithelial organoids; ex vivo culture; optical clearing; optical tissue clearing; CUBIC; SeeDB; 3DISCO; confocal; 3D imaging; advanced microscopy; advanced imaging; 3D confocal

Record Details Similar Records Cite « Share

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Harris, O. (2019). Elucidation of mammary gland development: combination of novel techniques to study proliferation and stem cells. (Doctoral Dissertation). University of Cambridge. Retrieved from https://www.repository.cam.ac.uk/handle/1810/291054

Chicago Manual of Style (16^th Edition):

Harris, Olivia. “Elucidation of mammary gland development: combination of novel techniques to study proliferation and stem cells.” 2019. Doctoral Dissertation, University of Cambridge. Accessed March 03, 2021. https://www.repository.cam.ac.uk/handle/1810/291054.

MLA Handbook (7^th Edition):

Harris, Olivia. “Elucidation of mammary gland development: combination of novel techniques to study proliferation and stem cells.” 2019. Web. 03 Mar 2021.

Vancouver:

Harris O. Elucidation of mammary gland development: combination of novel techniques to study proliferation and stem cells. [Internet] [Doctoral dissertation]. University of Cambridge; 2019. [cited 2021 Mar 03]. Available from: https://www.repository.cam.ac.uk/handle/1810/291054.

Council of Science Editors:

Harris O. Elucidation of mammary gland development: combination of novel techniques to study proliferation and stem cells. [Doctoral Dissertation]. University of Cambridge; 2019. Available from: https://www.repository.cam.ac.uk/handle/1810/291054

University of Cambridge

4. Harris, Olivia. Elucidation of mammary gland development : combination of novel techniques to study proliferation and stem cells.

Degree: PhD, 2019, University of Cambridge

URL: https://doi.org/10.17863/CAM.38235 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.774673

► The mammary gland is a dynamic organ that undergoes many cycles of proliferation and death throughout both oestrus cycling and the gestation/lactation/involution cycle. Moreover, it… (more)

▼ The mammary gland is a dynamic organ that undergoes many cycles of proliferation and death throughout both oestrus cycling and the gestation/lactation/involution cycle. Moreover, it is a unique tissue in that the majority of its development occurs postnatally coincident with the production of ovarian hormones in puberty. This capacity of the mammary gland for rapid growth and regeneration has been attributed to mammary stem cells (MaSCs). However, despite extensive efforts over the past 60 years, definitive characterisation of the stem and progenitor cells of the mammary gland has yet to be achieved. A number of recent conflicting, lineage tracing studies have served only to fuel the fires of controversy, with previous characterisation of the mammary gland largely carried out using two-dimensional tissue sections. However, in order to fully appreciate the capacity of MaSCs and to maintain spatial information of the complex topological structure, the mammary gland must be investigated in its intact form. Accordingly, there is still much disagreement regarding the potency, capacity and location of MaSCs. Consequently, in order to unequivocally elucidate the MaSC hierarchy, a variety of novel techniques have been combined in this thesis. The first involves development and optimisation of optical tissue clearing techniques to allow the visualisation of the native mammary gland, in situ, and in three dimensions. To do so, a number of different optical tissue clearing methods have been assessed and combined with a variety of microscopy techniques to allow the multiple focal planes of the ductal network to be examined. These imaging techniques were then combined with two neutral lineage tracing models; the first, the Rosa26[CA]30 model, utilises stochastic continuous clonal labelling to allow for the fate tracking of the progeny from single functional stem and progenitor cells. The second unbiased lineage tracing approach, the Rosa26-Confetti model, allows for the mammary stem and progenitor progeny to be traced with precise timing, with the additional benefit of a multicolour reporter. Next, proliferation was examined in wholemount tissues to investigate the functional requirements for MaSCs, and their potential locations. Finally, these techniques have been combined with an ex vivo 3D organoid culture system to investigate the use of culture methods in examining mammary epithelial cell dynamics. By combination of these techniques, clonally marked regions can be investigated throughout the development of the mammary gland, from the formation of the embryonic mammary iii rudiment, to expansion of the ductal tree in puberty, and ultimately their fate in lactation and involution, where the mammary gland fulfils its evolutionary purpose. The mammary gland provides a unique opportunity to investigate epithelial development extra-embryonically that is not available in other tissues. Moreover, study of maintenance and turnover of this organ has important implications for other epithelial systems. Finally, elucidation of the…

Subjects/Keywords: stem cells; mammary gland; adult stem cells; lineage tracing; proliferation; breast; breast cancer; MaSC; MaSCs; mammary gland stem cells; mammary stem cells; confetti; CA30; organoids; 3D models; mammary gland organoids; epithelial biology; epithelial organoids; ex vivo culture; optical clearing; optical tissue clearing; CUBIC; SeeDB; 3DISCO; confocal; 3D imaging; advanced microscopy; advanced imaging; 3D confocal

Record Details Similar Records Cite « Share

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Harris, O. (2019). Elucidation of mammary gland development : combination of novel techniques to study proliferation and stem cells. (Doctoral Dissertation). University of Cambridge. Retrieved from https://doi.org/10.17863/CAM.38235 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.774673

Chicago Manual of Style (16^th Edition):

Harris, Olivia. “Elucidation of mammary gland development : combination of novel techniques to study proliferation and stem cells.” 2019. Doctoral Dissertation, University of Cambridge. Accessed March 03, 2021. https://doi.org/10.17863/CAM.38235 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.774673.

MLA Handbook (7^th Edition):

Harris, Olivia. “Elucidation of mammary gland development : combination of novel techniques to study proliferation and stem cells.” 2019. Web. 03 Mar 2021.

Vancouver:

Harris O. Elucidation of mammary gland development : combination of novel techniques to study proliferation and stem cells. [Internet] [Doctoral dissertation]. University of Cambridge; 2019. [cited 2021 Mar 03]. Available from: https://doi.org/10.17863/CAM.38235 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.774673.

Council of Science Editors:

Harris O. Elucidation of mammary gland development : combination of novel techniques to study proliferation and stem cells. [Doctoral Dissertation]. University of Cambridge; 2019. Available from: https://doi.org/10.17863/CAM.38235 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.774673

Penn State University

5. Plichta, Kristin Ann. Mammary Epithelial Cell Subtype-Specific Analysis of Ras Pathway Activation.

Degree: 2010, Penn State University

URL: https://submit-etda.libraries.psu.edu/catalog/11251

► Breast cancers can be divided into subtypes based in part on how closely the tumor cells resemble mammary epithelial cell (MEC) subtypes resident in normal… (more)

▼ Breast cancers can be divided into subtypes based in part on how closely the tumor cells resemble mammary epithelial cell (MEC) subtypes resident in normal breast tissue. Though breast cancer subtypes differ in their aggressiveness and their response to treatment, the mechanisms by which distinct subtypes arise remain unknown. Traditionally, attempts to explain the varied clinical behavior of breast cancers rely on defining the genetic lesions harbored within individual tumors. However, recent evidence suggests that distinct breast cancer subtypes may arise from distinct MEC lineages, suggesting that some biological features of a given breast cancer subtype may be attributable to its antecedent “cell of origin”. Elucidating whether MEC lineage impacts the biology of descendant breast cancers presents a formidable challenge. By the time a tumor is clinically detectable, breast cancers have undergone clonal evolution that precludes a reliable retrospective determination of the cell of origin. As such, prospective analyses may be required to determine whether distinct MEC subtypes yield distinct cancer subtypes. We hypothesized that distinct MEC compartments yield different phenotypes in response to an identical oncogenic stimulus. To test this possibility, we generated mouse models that permit doxycycline-dependent expression of transgenes in an MEC compartment-restricted manner and developed strategies to monitor transgene-mediated phenotypes in real-time. As a first step toward studying malignant transformation of distinct MEC subtypes in mice, we tested experimental strategies designed to permit restriction of transgene expression to distinct MEC compartments in vitro. Transgenic mammary epithelium was partially disaggregated and propagated in 3D culture as mammary organoids, preserving the bilayered arrangement of the basal and luminal MEC compartments. Pairing a tet operator-driven H2B-eGFP reporter transgene with either a basal or luminal MEC transactivator enabled MEC compartment-restricted reporter gene expression. Notably, nuclear fluorescence resulting from H2B-eGFP expression enabled visualization of cellular dynamics within discrete MEC compartments. Through extended, multiparameter live cell imaging of organoids, time-lapse movies were generated that enabled visualization of MEC migration as well as tracking and quantitation of mitotic and apoptotic events. Next, we adapted this system to co-express the H2B-eGFP reporter together with a tet-operator-regulated oncogenic H-RASG12V allele. Whether expressed in a basal or luminal MEC-restricted manner, H-RASG12V expression reproducibly triggered aberrant organoid growth, reflected in measureable changes in organoid size and shape. Increases in organoid size were attributable to H-RASG12V-mediated increases in cell proliferation and decreases in cell death. In addition, H-RASG12V expression in either MEC compartment drove MECs to both traverse normal compartment boundaries and adopt modes of cellular migration distinct from those encountered in the setting… Advisors/Committee Members: Edward Joseph Gunther, Dissertation Advisor/Co-Advisor, Edward Joseph Gunther, Committee Chair/Co-Chair, Andrea Manni, Committee Member, Christopher Alan Siedlecki, Committee Member, Lisa M Shantz, Committee Member.

Subjects/Keywords: Ras; mammary organoids; breast cancer; transgenic mice; mammary epithelial cells

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Plichta, K. A. (2010). Mammary Epithelial Cell Subtype-Specific Analysis of Ras Pathway Activation. (Thesis). Penn State University. Retrieved from https://submit-etda.libraries.psu.edu/catalog/11251

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Plichta, Kristin Ann. “Mammary Epithelial Cell Subtype-Specific Analysis of Ras Pathway Activation.” 2010. Thesis, Penn State University. Accessed March 03, 2021. https://submit-etda.libraries.psu.edu/catalog/11251.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Plichta, Kristin Ann. “Mammary Epithelial Cell Subtype-Specific Analysis of Ras Pathway Activation.” 2010. Web. 03 Mar 2021.

Vancouver:

Plichta KA. Mammary Epithelial Cell Subtype-Specific Analysis of Ras Pathway Activation. [Internet] [Thesis]. Penn State University; 2010. [cited 2021 Mar 03]. Available from: https://submit-etda.libraries.psu.edu/catalog/11251.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Plichta KA. Mammary Epithelial Cell Subtype-Specific Analysis of Ras Pathway Activation. [Thesis]. Penn State University; 2010. Available from: https://submit-etda.libraries.psu.edu/catalog/11251

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Saskatchewan

6. Gilchrist, Samuel Edward. Transporter gene expression in rat lactating mammary epithelial cells & primary organoid cultures using quantitative real-time reverse transcription polymerase chain reaction (QRT-PCR).

Degree: 2007, University of Saskatchewan

URL: http://hdl.handle.net/10388/etd-01302007-094842

► Transporters dynamically expressed at the mammary gland transport critical nutrients into the breast milk of nursing mothers to meet the nutritional demands of the suckling… (more)

▼ Transporters dynamically expressed at the mammary gland transport critical nutrients into the breast milk of nursing mothers to meet the nutritional demands of the suckling infant. However, xenobiotics may interact with these transporters to potentially alter the nutrient composition of milk and compromise neonatal nutrition. The aim of the present study was to quantitatively evaluate the constitutive expression of various nutrient transporters in whole mammary gland tissue and mammary epithelial organoids (MEO) isolated from female Sprague-Dawley rats at various stages of pregnancy, lactation, and involution. Furthermore, the study’s aim was to determine if appropriately cultured mammary epithelial organoids (MEO) maintain in vivo transporter expression to lay down critical groundwork for the development of an in vitro screening tool assessing xenobiotic-nutrient transporter interactions. The following transporters were evaluated using quantitative real-time reverse transcription polymerase chain reaction (QRT-PCR): multidrug resistance protein (Mdr) 1a, 1b; multidrug resistance-like protein (Mrp) 1; organic cation transporter (Oct) 1; organic cation/carnitine transporter (Octn) 1, 2, and 3; concentrative nucleoside transporter (Cnt) 1, 2, and 3; equilibrative nucleoside transporter (Ent) 1, 2, and 3; nucleobase transporter (Ncbt) 1 and 2; oligopeptide transporter (Pept) 1 and 2; methotrexate carrier (Mtx) 1; divalent metal transporter (Dmt) 1; and the milk protein ?-casein. Transporter expression patterns in MEO differed from whole tissue for ?-actin, Mdr1a, Mdr1b, Oct1, Octn3, Ent3, Cnt1, Cnt3, Ncbt1, Pept2, Mtx1, and ?-casein. This brings into question whether whole mammary gland tissue is truly appropriate for an understanding of transporter expression in the mammary epithelium. Nevertheless, four general transporter expression patterns emerged in isolated MEO: decline throughout lactation (Mdr1a, Mdr1b, Mrp1 & Dmt1), increase throughout lactation (Cnt1 & Octn3), increase in early lactation (Oct1, Octn2, Ent1, Cnt2, Cnt3, Pept2 & Mtx1) and constant expression throughout lactation (Octn1, Ent2, Ent3, Ncbt1, Ncbt2 & Pept1). These expression patterns will provide insight into the critical windows of nutrient delivery to the breast milk to provide adequate nutritional stimuli to the suckling infant. Furthermore, MEO cultured in an extracellular matrix-rich environment maintained transporter expression at the mRNA level, which underscores the potential of the primary MEO in vitro model system as a screening tool for xenobiotic-transporter interactions at the mammary gland. Transporter expression patterns in MEO were unique for each transporter evaluated. This information accompanied by an in vitro screening tool may allow for predictions of xenobiotic interference with breast milk composition to help safeguard infant health. Advisors/Committee Members: Alcorn, Jane, Paterson, Phyllis G., Nazarali, Adil J., Maenz, David D..

Subjects/Keywords: real-time PCR; whole mammary tissue; mammary epithelial organoids; Mammary gland; transporters; lactation; rat

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Gilchrist, S. E. (2007). Transporter gene expression in rat lactating mammary epithelial cells & primary organoid cultures using quantitative real-time reverse transcription polymerase chain reaction (QRT-PCR). (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/etd-01302007-094842

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Gilchrist, Samuel Edward. “Transporter gene expression in rat lactating mammary epithelial cells & primary organoid cultures using quantitative real-time reverse transcription polymerase chain reaction (QRT-PCR).” 2007. Thesis, University of Saskatchewan. Accessed March 03, 2021. http://hdl.handle.net/10388/etd-01302007-094842.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Gilchrist, Samuel Edward. “Transporter gene expression in rat lactating mammary epithelial cells & primary organoid cultures using quantitative real-time reverse transcription polymerase chain reaction (QRT-PCR).” 2007. Web. 03 Mar 2021.

Vancouver:

Gilchrist SE. Transporter gene expression in rat lactating mammary epithelial cells & primary organoid cultures using quantitative real-time reverse transcription polymerase chain reaction (QRT-PCR). [Internet] [Thesis]. University of Saskatchewan; 2007. [cited 2021 Mar 03]. Available from: http://hdl.handle.net/10388/etd-01302007-094842.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Gilchrist SE. Transporter gene expression in rat lactating mammary epithelial cells & primary organoid cultures using quantitative real-time reverse transcription polymerase chain reaction (QRT-PCR). [Thesis]. University of Saskatchewan; 2007. Available from: http://hdl.handle.net/10388/etd-01302007-094842

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

7. Derecichei, Iulian Eric. Characterizing phenotypic effects of Spy1 mediated lateral branching.

Degree: MS, Biological Sciences, 2019, University of Windsor

URL: https://scholar.uwindsor.ca/etd/7693

► Mammary development is a continuous and cyclic process that is under tight regulatory control from hormones and cell cycle regulators to mediate transition from the… (more)

Subjects/Keywords: Biology; Cancer; Cell Biology; Mammary; Molecular Biology; Organoids

…23 Mammary Organoids, 3D culture and In-vitro models… …28 Generating Mammary Organoids… …1 The Mammary Gland… …1 Mouse Mammary Development in Embryogenesis and Puberty… …5 Molecular Signaling in Mammary Proliferation…

10 more images…

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Derecichei, I. E. (2019). Characterizing phenotypic effects of Spy1 mediated lateral branching. (Masters Thesis). University of Windsor. Retrieved from https://scholar.uwindsor.ca/etd/7693

Chicago Manual of Style (16^th Edition):

Derecichei, Iulian Eric. “Characterizing phenotypic effects of Spy1 mediated lateral branching.” 2019. Masters Thesis, University of Windsor. Accessed March 03, 2021. https://scholar.uwindsor.ca/etd/7693.

MLA Handbook (7^th Edition):

Derecichei, Iulian Eric. “Characterizing phenotypic effects of Spy1 mediated lateral branching.” 2019. Web. 03 Mar 2021.

Vancouver:

Derecichei IE. Characterizing phenotypic effects of Spy1 mediated lateral branching. [Internet] [Masters thesis]. University of Windsor; 2019. [cited 2021 Mar 03]. Available from: https://scholar.uwindsor.ca/etd/7693.

Council of Science Editors:

Derecichei IE. Characterizing phenotypic effects of Spy1 mediated lateral branching. [Masters Thesis]. University of Windsor; 2019. Available from: https://scholar.uwindsor.ca/etd/7693

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