title: Formation of microvascular networks in vitro.
abstract: This protocol describes how to form a 3D cell culture with explicit, endothelialized microvessels. The approach leads to fully enclosed, perfusable vessels in a bioremodelable hydrogel (type I collagen). The protocol uses microfabrication to enable user-defined geometries of the vascular network and microfluidic perfusion to control mass transfer and hemodynamic forces. These microvascular networks (μVNs) allow for multiweek cultures of endothelial cells or cocultures with parenchymal or tissue cells in the extra-lumen space. The platform enables real-time fluorescence imaging of living engineered tissues, in situ confocal fluorescence of fixed cultures and transmission electron microscopy (TEM) imaging of histological sections. This protocol enables studies of basic vascular and blood biology, provides a model for diseases such as tumor angiogenesis or thrombosis and serves as a starting point for constructing prevascularized tissues for regenerative medicine. After one-time microfabrication steps, the system can be assembled in less than 1 d and experiments can run for weeks.
authors.affiliation: School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, New York, USA.
authors.forename: John P
authors.initials: JP
authors.lastname: Morgan
authors.forename: Peter F
authors.initials: PF
authors.lastname: Delnero
authors.forename: Ying
authors.initials: Y
authors.lastname: Zheng
authors.forename: Scott S
authors.initials: SS
authors.lastname: Verbridge
authors.forename: Junmei
authors.initials: J
authors.lastname: Chen
authors.forename: Michael
authors.initials: M
authors.lastname: Craven
authors.forename: Nak Won
authors.initials: NW
authors.lastname: Choi
authors.forename: Anthony
authors.initials: A
authors.lastname: Diaz-Santana
authors.forename: Pouneh
authors.initials: P
authors.lastname: Kermani
authors.forename: Barbara
authors.initials: B
authors.lastname: Hempstead
authors.forename: José A
authors.initials: JA
authors.lastname: López
authors.forename: Thomas N
authors.initials: TN
authors.lastname: Corso
authors.forename: Claudia
authors.initials: C
authors.lastname: Fischbach
authors.forename: Abraham D
authors.initials: AD
authors.lastname: Stroock
mesh_terms: D018929:Cell Culture Techniques; D002478:Cells, Cultured; D018920:Coculture Techniques; D024042:Collagen Type I / Q000737:chemistry; D042783:Endothelial Cells; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D046529:Microscopy, Electron, Transmission; D055616:Microtechnology; D055806:Microvessels; D018919:Neovascularization, Physiologic; D061848:Optical Imaging; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods*
chemical_list: D024042:Collagen Type I; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate