In Vitro Modeling of the Neurovascular Environment by Coculturing Adult Human Brain Endothelial Cells with Human Neural Stem Cells
Identifieur interne : 000293 ( France/Analysis ); précédent : 000292; suivant : 000294In Vitro Modeling of the Neurovascular Environment by Coculturing Adult Human Brain Endothelial Cells with Human Neural Stem Cells
Auteurs : Chung-Hsing Chou [États-Unis, Royaume-Uni, Taïwan] ; John D. Sinden [Royaume-Uni] ; Pierre-Olivier Couraud [France] ; Michel Modo [États-Unis]Source :
- PLoS ONE [ 1932-6203 ] ; 2014.
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English descriptors
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Abstract
Brain and vascular cells form a functionally integrated signalling network that is known as the neurovascular unit (NVU). The signalling (autocrine, paracrine and juxtacrine) between different elements of this unit, especially in humans, is difficult to disentangle
Url:
DOI: 10.1371/journal.pone.0106346
PubMed: 25187991
PubMed Central: 4154686
Affiliations:
- France, Royaume-Uni, Taïwan, États-Unis
- Angleterre, Grand Londres, Pennsylvanie, Île-de-France
- Londres, Paris, Pittsburgh
- Université Paris-Descartes, Université de Pittsburgh
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PMC:4154686Le document en format XML
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wicri:level="4"><nlm:aff id="aff1"><addr-line>University of Pittsburgh, McGowan Institute for Regenerative Medicine, Pittsburgh, Pennsylvania, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>University of Pittsburgh, McGowan Institute for Regenerative Medicine, Pittsburgh, Pennsylvania</wicri:regionArea><placeName><region type="state">Pennsylvanie</region><settlement type="city">Pittsburgh</settlement></placeName><orgName type="university">Université de Pittsburgh</orgName></affiliation><affiliation wicri:level="4"><nlm:aff id="aff8"><addr-line>University of Pittsburgh, Department of Radiology, Pittsburgh, Pennsylvania, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>University of Pittsburgh, Department of Radiology, Pittsburgh, Pennsylvania</wicri:regionArea><placeName><region type="state">Pennsylvanie</region><settlement type="city">Pittsburgh</settlement></placeName><orgName type="university">Université de Pittsburgh</orgName></affiliation><affiliation wicri:level="4"><nlm:aff id="aff9"><addr-line>University of Pittsburgh, Department of Bioengineering, Pittsburgh, Pennsylvania, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>University of Pittsburgh, Department of Bioengineering, Pittsburgh, Pennsylvania</wicri:regionArea><placeName><region type="state">Pennsylvanie</region><settlement type="city">Pittsburgh</settlement></placeName><orgName type="university">Université de Pittsburgh</orgName></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Astrocytes (cytology)</term><term>Brain (cytology)</term><term>Cell Differentiation</term><term>Cell Line</term><term>Cells, Cultured</term><term>Coculture Techniques</term><term>Endothelial Cells (cytology)</term><term>Humans</term><term>Neural Stem Cells (cytology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Astrocytes (cytologie)</term><term>Cellules cultivées</term><term>Cellules endothéliales (cytologie)</term><term>Cellules souches neurales (cytologie)</term><term>Différenciation cellulaire</term><term>Encéphale (cytologie)</term><term>Humains</term><term>Lignée cellulaire</term><term>Techniques de coculture</term></keywords><keywords scheme="MESH" qualifier="cytologie" xml:lang="fr"><term>Astrocytes</term><term>Cellules endothéliales</term><term>Cellules souches neurales</term><term>Encéphale</term></keywords><keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Astrocytes</term><term>Brain</term><term>Endothelial Cells</term><term>Neural Stem Cells</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Cell Differentiation</term><term>Cell Line</term><term>Cells, Cultured</term><term>Coculture Techniques</term><term>Humans</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Cellules cultivées</term><term>Différenciation cellulaire</term><term>Humains</term><term>Lignée cellulaire</term><term>Techniques de coculture</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Brain and vascular cells form a functionally integrated signalling network that is known as the neurovascular unit (NVU). The signalling (autocrine, paracrine and juxtacrine) between different elements of this unit, especially in humans, is difficult to disentangle <italic>in vivo</italic>. Developing representative <italic>in vitro</italic> models is therefore essential to better understand the cellular interactions that govern the neurovascular environment. We here describe a novel approach to assay these cellular interactions by combining a human adult cerebral microvascular endothelial cell line (hCMEC/D3) with a fetal ganglionic eminence-derived neural stem cell (hNSC) line. These cell lines provide abundant homogeneous populations of cells to produce a consistently reproducible <italic>in vitro</italic> model of endothelial morphogenesis and the ensuing NVU. Vasculature-like structures (VLS) interspersed with patches of differentiating neural cells only occurred when hNSCs were seeded onto a differentiated endothelium. These VLS emerged within 3 days of coculture and by day 6 were stabilizing. After 7 days of coculture, neuronal differentiation of hNSCs was increased 3-fold, but had no significant effect on astrocyte or oligodendrocyte differentiation. ZO1, a marker of adherens and tight junctions, was highly expressed in both undifferentiated and differentiated endothelial cells, but the adherens junction markers CD31 and VE-cadherin were significantly reduced in coculture by approximately 20%. A basement membrane, consisting of laminin, vitronectin, and collagen I and IV, separated the VLS from neural patches. This simple assay can assist in elucidating the cellular and molecular signaling involved in the formation of VLS, as well as the enhancement of neuronal differentiation through endothelial signaling.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Hawkins, Bt" uniqKey="Hawkins B">BT Hawkins</name></author><author><name sortKey="Davis, Tp" uniqKey="Davis T">TP Davis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kleinfeld, D" uniqKey="Kleinfeld D">D Kleinfeld</name></author><author><name sortKey="Blinder, P" uniqKey="Blinder P">P Blinder</name></author><author><name sortKey="Drew, Pj" uniqKey="Drew P">PJ Drew</name></author><author><name sortKey="Driscoll, Jd" uniqKey="Driscoll J">JD Driscoll</name></author><author><name sortKey="Muller, A" uniqKey="Muller A">A Muller</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Moeller, Bj" uniqKey="Moeller B">BJ 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S" uniqKey="Liu S">S Liu</name></author><author><name sortKey="Agalliu, D" uniqKey="Agalliu D">D Agalliu</name></author><author><name sortKey="Yu, C" uniqKey="Yu C">C Yu</name></author><author><name sortKey="Fisher, M" uniqKey="Fisher M">M Fisher</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Takeshita, Y" uniqKey="Takeshita Y">Y Takeshita</name></author><author><name sortKey="Ransohoff, Rm" uniqKey="Ransohoff R">RM Ransohoff</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Arnold, T" uniqKey="Arnold T">T Arnold</name></author><author><name sortKey="Betsholtz, C" uniqKey="Betsholtz C">C Betsholtz</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Baker, Bm" uniqKey="Baker B">BM Baker</name></author><author><name sortKey="Chen, Cs" uniqKey="Chen C">CS Chen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Eng, G" uniqKey="Eng G">G Eng</name></author><author><name sortKey="Lee, Bw" uniqKey="Lee B">BW Lee</name></author><author><name sortKey="Parsa, H" uniqKey="Parsa H">H Parsa</name></author><author><name sortKey="Chin, Cd" uniqKey="Chin C">CD Chin</name></author><author><name sortKey="Schneider, J" uniqKey="Schneider J">J Schneider</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>France</li><li>Royaume-Uni</li><li>Taïwan</li><li>États-Unis</li></country><region><li>Angleterre</li><li>Grand Londres</li><li>Pennsylvanie</li><li>Île-de-France</li></region><settlement><li>Londres</li><li>Paris</li><li>Pittsburgh</li></settlement><orgName><li>Université Paris-Descartes</li><li>Université de Pittsburgh</li></orgName></list><tree><country name="États-Unis"><region name="Pennsylvanie"><name sortKey="Chou, Chung Hsing" sort="Chou, Chung Hsing" uniqKey="Chou C" first="Chung-Hsing" last="Chou">Chung-Hsing Chou</name></region><name sortKey="Modo, Michel" sort="Modo, Michel" uniqKey="Modo M" first="Michel" last="Modo">Michel Modo</name><name sortKey="Modo, Michel" sort="Modo, Michel" uniqKey="Modo M" first="Michel" last="Modo">Michel Modo</name><name sortKey="Modo, Michel" sort="Modo, Michel" uniqKey="Modo M" first="Michel" last="Modo">Michel Modo</name></country><country name="Royaume-Uni"><region name="Angleterre"><name sortKey="Chou, Chung Hsing" sort="Chou, Chung Hsing" uniqKey="Chou C" first="Chung-Hsing" last="Chou">Chung-Hsing Chou</name></region><name sortKey="Sinden, John D" sort="Sinden, John D" uniqKey="Sinden J" first="John D." last="Sinden">John D. Sinden</name></country><country name="Taïwan"><noRegion><name sortKey="Chou, Chung Hsing" sort="Chou, Chung Hsing" uniqKey="Chou C" first="Chung-Hsing" last="Chou">Chung-Hsing Chou</name></noRegion></country><country name="France"><region name="Île-de-France"><name sortKey="Couraud, Pierre Olivier" sort="Couraud, Pierre Olivier" uniqKey="Couraud P" first="Pierre-Olivier" last="Couraud">Pierre-Olivier Couraud</name></region><name sortKey="Couraud, Pierre Olivier" sort="Couraud, Pierre Olivier" uniqKey="Couraud P" first="Pierre-Olivier" last="Couraud">Pierre-Olivier Couraud</name><name sortKey="Couraud, Pierre Olivier" sort="Couraud, Pierre Olivier" uniqKey="Couraud P" first="Pierre-Olivier" last="Couraud">Pierre-Olivier Couraud</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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