Formation of microvascular networks in vitro

Nat Protoc. 2013 Sep;8(9):1820-36. doi: 10.1038/nprot.2013.110. Epub 2013 Aug 29.


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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Cell Culture Techniques
  • Cells, Cultured
  • Coculture Techniques
  • Collagen Type I / chemistry
  • Endothelial Cells
  • Humans
  • Hydrogel, Polyethylene Glycol Dimethacrylate / chemistry
  • Microscopy, Electron, Transmission
  • Microtechnology
  • Microvessels*
  • Neovascularization, Physiologic
  • Optical Imaging
  • Tissue Engineering / instrumentation
  • Tissue Engineering / methods*


  • Collagen Type I
  • Hydrogel, Polyethylene Glycol Dimethacrylate