Isolation, differentiation and characterization of vascular cells derived from human embryonic stem cells

• Main Authors: Levenberg, Shulamit (Author), Ferreira, Lino S. (Author), Chen-Konak, Limor (Author), Kraehenbuehl, Thomas P (Author), Langer, Robert S (Author)
• Other Authors: Massachusetts Institute of Technology. Department of Chemical Engineering (Contributor)
• Format: Article
• Language: English
• Published: Springer Nature America, Inc, 2020-08-13T22:50:42Z.
• Subjects: Article
• Online Access: Get fulltext

 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