Journal
NATURE PROTOCOLS
Volume 8, Issue 9, Pages 1820-1836Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2013.110
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Funding
- American Heart Association Scientist Development Grant
- US National Institutes of Health (NIH) [R01HL091153, RC1 CA146065]
- Cornell Center on the Microenvironment and Metastasis [NCI-U54 CA143876]
- Human Frontiers in Science Program
- Cornell Nanobiotechnology Center [NSF-STC, ECS-9876771]
- Cornell Center for Nanoscale Science and Technology [NSF-NNIN ECS 03-35765]
- Empire State Development Division of Science, Technology and Innovation (NYSTAR) Center for Advanced Technology (CAT) award
- New York State J.D. Watson Award
- Arnold and Mabel Beckman Foundation Young Investigator Award
- National Science Foundation Graduate Fellowship
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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.
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