title: In vitro self-assembly of human pericyte-supported endothelial microvessels in three-dimensional coculture: a simple model for interrogating endothelial-pericyte interactions.
abstract: We describe a method for coculture 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 antiapoptotic protein Bcl-2. To prevent PC-mediated gel contraction, the collagen-fibronectin gel was polymerized within a polyglycolic acid nonwoven 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 cocultures, 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.
authors.affiliation: Department of Immunobiology, Yale University School of Medicine, New Haven, CT, USA. jpw55@cam.ac.uk
authors.forename: J P
authors.initials: JP
authors.lastname: Waters
authors.forename: M S
authors.initials: MS
authors.lastname: Kluger
authors.forename: M
authors.initials: M
authors.lastname: Graham
authors.forename: W G
authors.initials: WG
authors.lastname: Chang
authors.forename: J R
authors.initials: JR
authors.lastname: Bradley
authors.forename: J S
authors.initials: JS
authors.lastname: Pober
mesh_terms: D017209:Apoptosis; D002450:Cell Communication; D002465:Cell Movement; D002478:Cells, Cultured; D018920:Coculture Techniques; D003094:Collagen / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism / Q000502:physiology*; D005353:Fibronectins / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D007365:Intercellular Junctions / Q000502:physiology; D055806:Microvessels / Q000378:metabolism / Q000502:physiology*; D018919:Neovascularization, Physiologic; D020286:Pericytes / Q000378:metabolism / Q000502:physiology*; D011100:Polyglycolic Acid / Q000737:chemistry; D058105:Polymerization; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014161:Transduction, Genetic; D014162:Transfection
chemical_list: D005353:Fibronectins; D019253:Proto-Oncogene Proteins c-bcl-2; D011100:Polyglycolic Acid; D003094:Collagen