Morgan, John P. Delnero, Peter F. Zheng, Ying Verbridge, Scott S. Chen, Junmei Craven, Michael Choi, Nak Won Diaz-Santana, Anthony Kermani, Pouneh Hempstead, Barbara Lopez, Jose A. Corso, Thomas N. Fischbach, Claudia Stroock, Abraham D. 2024-01-20T11:34:21Z 2024-01-20T11:34:21Z 2021-08-31 2013-09 1754-2189 https://pubs.kist.re.kr/handle/201004/127747 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 (mVNs) 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. English NATURE PUBLISHING GROUP EXTRACELLULAR MATRICES VASCULARIZED TISSUE SOFT LITHOGRAPHY ANGIOGENESIS CULTURE MORPHOGENESIS MICROVESSELS FABRICATION PRINCIPLES HYDROGELS Formation of microvascular networks in vitro Article 10.1038/nprot.2013.110 1 NATURE PROTOCOLS, v.8, no.9, pp.1820 - 1836 NATURE PROTOCOLS 8 9 1820 1836 scie scopus 000323768600015 2-s2.0-84883342307 Biochemical Research Methods Biochemistry & Molecular Biology Article EXTRACELLULAR MATRICES VASCULARIZED TISSUE SOFT LITHOGRAPHY ANGIOGENESIS CULTURE MORPHOGENESIS MICROVESSELS FABRICATION PRINCIPLES HYDROGELS