Journal
NATURE PROTOCOLS
Volume 5, Issue 6, Pages 1115-1126Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2010.31
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Funding
- NIH [HL060435, DE013023]
- Marie-Curie Reintegration Grants
- MIT-Portugal
- Crioestaminal/Associacao Viver a Ciencia
- FCT [PTDC/SAU-BEB/098468/2008]
- Swiss National Science Foundation [PBELP3-127902]
- NATIONAL HEART, LUNG, AND BLOOD INSTITUTE [R01HL060435] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH [R01DE013023] Funding Source: NIH RePORTER
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Herein, we describe a protocol for the isolation of human embryonic stem cells (hESCs)-derived vascular cells at various stages of development. the cells are isolated from 10 to 15-d-old human embryoid bodies (EBs) cultured in suspension. after dissociation, cells are labeled with anti-CD34 or anti-CD31 (PECAM1) antibody and separated from the cell mixture by magnetic-activated cell separation (Macs) or fluorescent-activated cell sorting (FACS). Isolated vascular cells are then cultured in media conditions that support specific differentiation and expansion pathways. the resulting vascular cell populations contain >80% endothelial-like or smooth muscle-like cells. assuming typical initial cell adhesion and proliferation rates, the entire procedure can be completed within 1.5 months. Vascular cells isolated and differentiated under the described conditions may constitute a potential cell source for therapeutic application toward repair of ischemic tissues, preparation of tissue-engineered vascular grafts and design of cellular kits for drug screening applications.
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