Advanced search options

Advanced Search Options 🞨

Browse by author name (“Author name starts with…”).

Find ETDs with:

in
/  
in
/  
in
/  
in

Written in Published in Earliest date Latest date

Sorted by

Results per page:

You searched for id:"handle:10616/45513". One record found.

Search Limiters

Last 2 Years | English Only

No search limiters apply to these results.

▼ Search Limiters

1. Nordin, Joel Z. Purification and biodistribution of extracellular vesicles .

Degree: 2017, Karolinska Institute

Extracellular vesicles (EVs) are nano-sized vesicles that contain bioactive lipids, RNAs and proteins, which can be transferred to recipient cells. EVs are important for physiological as well as pathological processes, such as coagulation and immune homeostasis, aiding cancer metastasis and spread of infectious diseases. Owing to their relatively small size the purification of EVs is a challenge, hence we have established and optimised workflows consisting of ultrafiltration with subsequent size exclusion liquid chromatography (UFLC)( Paper I) and bind-elute combined with size exclusion (BE-SEC) columns (Paper III) for EV purification. UF-LC allowed for purification of biophysically intact EVs with better yield and purity compared to ultracentrifugation (UC), which is the gold standard purification method in the field. The biodistribution of UF-LC EVs was different compared to vesicles isolated using UC, despite having highly similar protein composition according to proteomics analysis. We found that UF-LC vesicles accumulated less in lung, possibly owing to their higher integrity. Indeed, fluorescence correlation spectroscopy and transmission electron microscopy indicated that the high gravitational forces in UC lead to aggregation and disruption of the vesicles. The BE-SEC method is a similar method to UF-LC, however protein impurities less than 700 kDa in size are bound in the interior of the beads, thus improving simple size-based exclusion. The BE-SEC method is scalable, produces samples with better purity than UC, displaying yields exceeding 70% and demonstrates a good reproducibility between samples. Moreover, vesicles purified by BE-SEC display the same EV surface markers as UC purified EVs, and CD63-eGFP positive vesicles are taken up in recipient cells to the same extent. In summary, the BE-SEC method is a reproducible and fast alternative to UF-LC for large media volumes. Reliable purification methods are important for the implementation of therapeutically active EVs, however knowledge regarding their eventual organotropism and biodistribution is equally important. Thus, in article II we evaluated the biodistribution of EVs specifically labeled with a near-infrared dye. The main sites of accumulation of exogenously injected EVs were liver, spleen and lungs. Biodistribution profile of EVs depended strongly on injection route, and to certain extent, on EV cell type source, as dendritic cell derived EVs exhibited a more pronounced uptake in spleen compared to the other cell sources tested. We further showed that small alterations of EV surface proteins could significantly affect biodistribution as well, since EVs equipped with a brain targeting peptide on their surface increased the uptake of targeted EVs in brain. This study highlights that the biodistribution of EVs follows other nano-sized particles with uptake mainly in liver. Administration route, cell source and a targeting peptide influence the distribution, however the overall distribution is unaltered…

Record DetailsSimilar RecordsGoogle PlusoneFacebookTwitterCiteULikeMendeleyreddit

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

APA (6th Edition):

Nordin, J. Z. (2017). Purification and biodistribution of extracellular vesicles . (Thesis). Karolinska Institute. Retrieved from http://hdl.handle.net/10616/45513

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):

Nordin, Joel Z. “Purification and biodistribution of extracellular vesicles .” 2017. Thesis, Karolinska Institute. Accessed December 18, 2017. http://hdl.handle.net/10616/45513.

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

MLA Handbook (7th Edition):

Nordin, Joel Z. “Purification and biodistribution of extracellular vesicles .” 2017. Web. 18 Dec 2017.

Vancouver:

Nordin JZ. Purification and biodistribution of extracellular vesicles . [Internet] [Thesis]. Karolinska Institute; 2017. [cited 2017 Dec 18]. Available from: http://hdl.handle.net/10616/45513.

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

Council of Science Editors:

Nordin JZ. Purification and biodistribution of extracellular vesicles . [Thesis]. Karolinska Institute; 2017. Available from: http://hdl.handle.net/10616/45513

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

.