In vitro self-assembly of human pericyte-supported endothelial microvessels in three-dimensional co-culture: a simple model for interrogating endothelial:pericyte interactions
Identifieur interne : 003921 ( Pmc/Curation ); précédent : 003920; suivant : 003922In vitro self-assembly of human pericyte-supported endothelial microvessels in three-dimensional co-culture: a simple model for interrogating endothelial:pericyte interactions
Auteurs : Jp Waters [Royaume-Uni] ; Ms Kluger [États-Unis] ; M. Graham [États-Unis] ; Wg Chang [États-Unis] ; Jr Bradley [Royaume-Uni] ; Js Pober [États-Unis]Source :
- Journal of vascular research [ 1018-1172 ] ; 2013.
Abstract
We describe a method for co-culture of macro or microvascular human endothelial cells (ECs) and pericytes (PCs) within a 3-dimensional (3-D) protein matrix resulting in lumenized EC cords invested by PCs. To prevent apoptotic cell death of ECs in 3-D culture, human umbilical vein or dermal microvascular ECs were transduced to express the anti-apoptotic protein Bcl-2. To prevent PC-mediated gel contraction, the collagen-fibronectin gel was polymerized within a polyglycolic acid (PGA) non-woven matrix. Over the first 24-48 h, EC-only gels spontaneously formed cords that developed lumens via vacuolization; such vascular networks were maintained for up to 7 days. In EC-PC co-cultures, PCs were recruited to the EC networks. PC investment of EC cords both limited the lumen diameter and increased the degree of vascular network arborization. Peg and socket junctions formed between ECs and PCs in this system but dye transfer, indicative of gap junction formation, was not observed. This simple system can be used to analyze bidirectional signals between ECs and PCs in a 3-D geometry.
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DOI: 10.1159/000353303
PubMed: 23860328
PubMed Central: 3879598
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self-assembly of human pericyte-supported endothelial microvessels in three-dimensional co-culture: a simple model for interrogating endothelial:pericyte interactions</title>
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self-assembly of human pericyte-supported endothelial microvessels in three-dimensional co-culture: a simple model for interrogating endothelial:pericyte interactions</title>
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<front><div type="abstract" xml:lang="en"><p id="P1">We describe a method for co-culture of macro or microvascular human endothelial cells (ECs) and pericytes (PCs) within a 3-dimensional (3-D) protein matrix resulting in lumenized EC cords invested by PCs. To prevent apoptotic cell death of ECs in 3-D culture, human umbilical vein or dermal microvascular ECs were transduced to express the anti-apoptotic protein Bcl-2. To prevent PC-mediated gel contraction, the collagen-fibronectin gel was polymerized within a polyglycolic acid (PGA) non-woven matrix. Over the first 24-48 h, EC-only gels spontaneously formed cords that developed lumens via vacuolization; such vascular networks were maintained for up to 7 days. In EC-PC co-cultures, PCs were recruited to the EC networks. PC investment of EC cords both limited the lumen diameter and increased the degree of vascular network arborization. Peg and socket junctions formed between ECs and PCs in this system but dye transfer, indicative of gap junction formation, was not observed. This simple system can be used to analyze bidirectional signals between ECs and PCs in a 3-D geometry.</p>
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self-assembly of human pericyte-supported endothelial microvessels in three-dimensional co-culture: a simple model for interrogating endothelial:pericyte interactions</article-title>
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<aff id="A1"><label>1</label>
Department of Immunobiology, Yale University School of Medicine, New Haven, Connecticut 06520, USA</aff>
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Center for Cellular and Molecular Imaging, Yale University School of Medicine, New Haven, Connecticut 06520, USA</aff>
<aff id="A3"><label>3</label>
Department of Medicine, University of Cambridge, Addenbrooke’s Hospital, Hills Road, Cambridge, CB2 0QQ, UK</aff>
<author-notes><corresp id="cor1"><bold>ADRESS OF CORRESPONDING AUTHOR</bold>
John P Waters, Department of Medicine, University Of Cambridge, Level 5 (Box 157), Addenbrooke’s Hospital, Hills Road, Cambridge, CB2 0QQ, UK</corresp>
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<abstract><p id="P1">We describe a method for co-culture of macro or microvascular human endothelial cells (ECs) and pericytes (PCs) within a 3-dimensional (3-D) protein matrix resulting in lumenized EC cords invested by PCs. To prevent apoptotic cell death of ECs in 3-D culture, human umbilical vein or dermal microvascular ECs were transduced to express the anti-apoptotic protein Bcl-2. To prevent PC-mediated gel contraction, the collagen-fibronectin gel was polymerized within a polyglycolic acid (PGA) non-woven matrix. Over the first 24-48 h, EC-only gels spontaneously formed cords that developed lumens via vacuolization; such vascular networks were maintained for up to 7 days. In EC-PC co-cultures, PCs were recruited to the EC networks. PC investment of EC cords both limited the lumen diameter and increased the degree of vascular network arborization. Peg and socket junctions formed between ECs and PCs in this system but dye transfer, indicative of gap junction formation, was not observed. This simple system can be used to analyze bidirectional signals between ECs and PCs in a 3-D geometry.</p>
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<kwd-group><kwd>Vasculogensis</kwd>
<kwd>3-Dimensional</kwd>
<kwd>Pericyte</kwd>
<kwd>Endothelial</kwd>
<kwd>Co-culture</kwd>
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