Isolation, differentiation and characterization of vascular cells derived from human embryonic stem cells
Author(s)
Levenberg, Shulamit; Ferreira, Lino S.; Chen-Konak, Limor; Kraehenbuehl, Thomas P; Langer, Robert S
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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. ©2010
Date issued
2010-05Department
Massachusetts Institute of Technology. Department of Chemical EngineeringJournal
Nature Protocols
Publisher
Springer Nature America, Inc
Citation
Levenberg, Shulamit et al., "Isolation, differentiation and characterization of vascular cells derived from human embryonic stem cells." Nature Protocols 5, 6 (June 2010): p. 1115–26 doi. 10.1038/nprot.2010.31 ©2010 Authors
Version: Author's final manuscript
ISSN
1750-2799