Generation, expansion and functional analysis of endothelial cells and pericytes derived from human pluripotent stem cells
The authors of this Protocol describe how to generate endothelial cells and pericytes and how to functionally evaluate the cells' ability to generate primary vascular plexus and incorporate it into the zebrafish vasculature. Human endothelial cells (ECs) and pericytes are of great interest for...
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| Veröffentlicht in: | Nature protocols 2014-06, Vol.9 (6), p.1514-1531 |
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| creator | Orlova, Valeria V van den Hil, Francijna E Petrus-Reurer, Sandra Drabsch, Yvette ten Dijke, Peter Mummery, Christine L |
| description | The authors of this Protocol describe how to generate endothelial cells and pericytes and how to functionally evaluate the cells' ability to generate primary vascular plexus and incorporate it into the zebrafish vasculature.
Human endothelial cells (ECs) and pericytes are of great interest for research on vascular development and disease, as well as for future therapy. This protocol describes the efficient generation of ECs and pericytes from human pluripotent stem cells (hPSCs) under defined conditions. Essential steps for hPSC culture, differentiation, isolation and functional characterization of ECs and pericytes are described. Substantial numbers of both cell types can be derived in only 2–3 weeks: this involves differentiation (10 d), isolation (1 d) and 4 or 10 d of expansion of ECs and pericytes, respectively. We also describe two assays for functional evaluation of hPSC-derived ECs: (i) primary vascular plexus formation upon coculture with hPSC-derived pericytes and (ii) incorporation in the vasculature of zebrafish xenografts
in vivo
. These assays can be used to test the quality and drug sensitivity of hPSC-derived ECs and model vascular diseases with patient-derived hPSCs. |
| doi_str_mv | 10.1038/nprot.2014.102 |
| format | Article |
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Human endothelial cells (ECs) and pericytes are of great interest for research on vascular development and disease, as well as for future therapy. This protocol describes the efficient generation of ECs and pericytes from human pluripotent stem cells (hPSCs) under defined conditions. Essential steps for hPSC culture, differentiation, isolation and functional characterization of ECs and pericytes are described. Substantial numbers of both cell types can be derived in only 2–3 weeks: this involves differentiation (10 d), isolation (1 d) and 4 or 10 d of expansion of ECs and pericytes, respectively. We also describe two assays for functional evaluation of hPSC-derived ECs: (i) primary vascular plexus formation upon coculture with hPSC-derived pericytes and (ii) incorporation in the vasculature of zebrafish xenografts
in vivo
. These assays can be used to test the quality and drug sensitivity of hPSC-derived ECs and model vascular diseases with patient-derived hPSCs.</description><identifier>ISSN: 1754-2189</identifier><identifier>EISSN: 1750-2799</identifier><identifier>DOI: 10.1038/nprot.2014.102</identifier><identifier>PMID: 24874816</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>631/1647/1407/651 ; 631/1647/334/1874/763 ; 631/443/1338 ; 631/532/1360 ; Analytical Chemistry ; Animals ; Biological Techniques ; Blood vessels ; Cell culture ; Cell Culture Techniques - methods ; Cell differentiation ; Cell Differentiation - physiology ; Cell Proliferation ; Composition ; Computational Biology/Bioinformatics ; Connective tissue cells ; Cryopreservation ; Danio rerio ; Differentiation ; Disease ; Endothelial cells ; Endothelial Cells - cytology ; Endothelial Cells - physiology ; Endothelium ; Flow cytometry ; Functional analysis ; Health aspects ; Heterografts - blood supply ; Heterografts - cytology ; Humans ; In vivo methods and tests ; Life Sciences ; Medical research ; Medicine, Experimental ; Methods ; Microarrays ; Organic Chemistry ; Pericytes ; Pericytes - cytology ; Pericytes - physiology ; Physiological aspects ; Pluripotency ; Pluripotent Stem Cells - cytology ; Pluripotent Stem Cells - physiology ; Protocol ; Stem cells ; Vascular diseases ; Xenografts ; Xenotransplantation ; Zebrafish</subject><ispartof>Nature protocols, 2014-06, Vol.9 (6), p.1514-1531</ispartof><rights>Springer Nature Limited 2014</rights><rights>COPYRIGHT 2014 Nature Publishing Group</rights><rights>Copyright Nature Publishing Group Jun 2014</rights><rights>Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. 2014.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c558t-bc5598883092c75c2e48db25e1edee073301334aabfbd3621bd134adda0587a03</citedby><cites>FETCH-LOGICAL-c558t-bc5598883092c75c2e48db25e1edee073301334aabfbd3621bd134adda0587a03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/nprot.2014.102$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/nprot.2014.102$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24874816$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Orlova, Valeria V</creatorcontrib><creatorcontrib>van den Hil, Francijna E</creatorcontrib><creatorcontrib>Petrus-Reurer, Sandra</creatorcontrib><creatorcontrib>Drabsch, Yvette</creatorcontrib><creatorcontrib>ten Dijke, Peter</creatorcontrib><creatorcontrib>Mummery, Christine L</creatorcontrib><title>Generation, expansion and functional analysis of endothelial cells and pericytes derived from human pluripotent stem cells</title><title>Nature protocols</title><addtitle>Nat Protoc</addtitle><addtitle>Nat Protoc</addtitle><description>The authors of this Protocol describe how to generate endothelial cells and pericytes and how to functionally evaluate the cells' ability to generate primary vascular plexus and incorporate it into the zebrafish vasculature.
Human endothelial cells (ECs) and pericytes are of great interest for research on vascular development and disease, as well as for future therapy. This protocol describes the efficient generation of ECs and pericytes from human pluripotent stem cells (hPSCs) under defined conditions. Essential steps for hPSC culture, differentiation, isolation and functional characterization of ECs and pericytes are described. Substantial numbers of both cell types can be derived in only 2–3 weeks: this involves differentiation (10 d), isolation (1 d) and 4 or 10 d of expansion of ECs and pericytes, respectively. We also describe two assays for functional evaluation of hPSC-derived ECs: (i) primary vascular plexus formation upon coculture with hPSC-derived pericytes and (ii) incorporation in the vasculature of zebrafish xenografts
in vivo
. 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Academic</collection><jtitle>Nature protocols</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Orlova, Valeria V</au><au>van den Hil, Francijna E</au><au>Petrus-Reurer, Sandra</au><au>Drabsch, Yvette</au><au>ten Dijke, Peter</au><au>Mummery, Christine L</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Generation, expansion and functional analysis of endothelial cells and pericytes derived from human pluripotent stem cells</atitle><jtitle>Nature protocols</jtitle><stitle>Nat Protoc</stitle><addtitle>Nat Protoc</addtitle><date>2014-06-01</date><risdate>2014</risdate><volume>9</volume><issue>6</issue><spage>1514</spage><epage>1531</epage><pages>1514-1531</pages><issn>1754-2189</issn><eissn>1750-2799</eissn><abstract>The authors of this Protocol describe how to generate endothelial cells and pericytes and how to functionally evaluate the cells' ability to generate primary vascular plexus and incorporate it into the zebrafish vasculature.
Human endothelial cells (ECs) and pericytes are of great interest for research on vascular development and disease, as well as for future therapy. This protocol describes the efficient generation of ECs and pericytes from human pluripotent stem cells (hPSCs) under defined conditions. Essential steps for hPSC culture, differentiation, isolation and functional characterization of ECs and pericytes are described. Substantial numbers of both cell types can be derived in only 2–3 weeks: this involves differentiation (10 d), isolation (1 d) and 4 or 10 d of expansion of ECs and pericytes, respectively. We also describe two assays for functional evaluation of hPSC-derived ECs: (i) primary vascular plexus formation upon coculture with hPSC-derived pericytes and (ii) incorporation in the vasculature of zebrafish xenografts
in vivo
. These assays can be used to test the quality and drug sensitivity of hPSC-derived ECs and model vascular diseases with patient-derived hPSCs.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>24874816</pmid><doi>10.1038/nprot.2014.102</doi><tpages>18</tpages></addata></record> |
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| subjects | 631/1647/1407/651 631/1647/334/1874/763 631/443/1338 631/532/1360 Analytical Chemistry Animals Biological Techniques Blood vessels Cell culture Cell Culture Techniques - methods Cell differentiation Cell Differentiation - physiology Cell Proliferation Composition Computational Biology/Bioinformatics Connective tissue cells Cryopreservation Danio rerio Differentiation Disease Endothelial cells Endothelial Cells - cytology Endothelial Cells - physiology Endothelium Flow cytometry Functional analysis Health aspects Heterografts - blood supply Heterografts - cytology Humans In vivo methods and tests Life Sciences Medical research Medicine, Experimental Methods Microarrays Organic Chemistry Pericytes Pericytes - cytology Pericytes - physiology Physiological aspects Pluripotency Pluripotent Stem Cells - cytology Pluripotent Stem Cells - physiology Protocol Stem cells Vascular diseases Xenografts Xenotransplantation Zebrafish |
| title | Generation, expansion and functional analysis of endothelial cells and pericytes derived from human pluripotent stem cells |
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