Differentiation of Endothelial Cells from Human Embryonic Stem Cells and Induced Pluripotent Stem Cells

  • Shijun Hu
  • Preston Lavinghousez
  • Zongjin Li
  • Joseph C. WuEmail author
Part of the Springer Protocols Handbooks book series (SPH)


Endothelial cells line the entire circulatory system and form the interface between the blood vessel intima and the circulating red blood cells. Endothelial cells are crucial to the proper function of the circulatory system and tissue viability, including their roles in coagulation, fibrinolysis, inflammation, and most specifically, vasculogenesis and angiogenesis. Given the importance of endothelial cells in vascular formation, it is essential to expand our knowledge of endothelial cell physiology and growth. The vasculogenic and angiogenic properties that enable new vascular networks to form and consequently perfuse ischemic tissues make endothelial cells an essential element of potential novel therapies. In this chapter, we describe a three-step technique to derive endothelial cells from human embryonic stem cells or induced pluripotent stem cells using a three-dimensional embryoid body formation protocol. A technique to derive a highly pure endothelial population using flow cytometry will also be discussed.

Key words

Human embryonic stem cells Induced pluripotent stem cells Endothelial cells Embryoid body Angiogenesis Vasculogenesis 


  1. 1.
    Thomson, J.A., et al., Embryonic stem cell lines derived from human blastocysts. Science, 1998. 282(5391): p. 1145–7.PubMedCrossRefGoogle Scholar
  2. 2.
    Takahashi, K., et al., Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell, 2007. 131(5): p. 861–72.PubMedCrossRefGoogle Scholar
  3. 3.
    Yu, J., et al., Induced pluripotent stem cell lines derived from human somatic cells. Science, 2007. 318(5858): p. 1917–20.PubMedCrossRefGoogle Scholar
  4. 4.
    Sun, N., M.T. Longaker, and J.C. Wu, Human iPS cell-based therapy: considerations before clinical applications. Cell Cycle, 2010. 9(5): p. 880–5.PubMedCrossRefGoogle Scholar
  5. 5.
    Kocher, A.A., et al., Neovascularization of ischemic myocardium by human bone-­marrow-derived angioblasts prevents cardiomyocyte apoptosis, reduces remodeling and improves cardiac function. Nat Med, 2001. 7(4): p. 430–6.PubMedCrossRefGoogle Scholar
  6. 6.
    Li, Z., et al., Differentiation, survival, and function of embryonic stem cell derived endothelial cells for ischemic heart disease. Circulation, 2007. 116(11 Suppl): p. I46-54.PubMedGoogle Scholar
  7. 7.
    Yu, J., et al., nAChRs mediate human embryonic stem cell-derived endothelial cells: proliferation, apoptosis, and angiogenesis. PLoS One, 2009. 4(9): p. e7040.PubMedCrossRefGoogle Scholar
  8. 8.
    Kooreman, N.G. and J.C. Wu, Tumorigenicity of pluripotent stem cells: biological insights from molecular imaging. J R Soc Interface, 2010.Google Scholar
  9. 9.
    Li, Z., Z. Han, and J.C. Wu, Transplantation of human embryonic stem cell-derived endothelial cells for vascular diseases. J Cell Biochem, 2009. 106(2): p. 194–9.PubMedCrossRefGoogle Scholar
  10. 10.
    Wang, Z.Z., et al., Endothelial cells derived from human embryonic stem cells form durable blood vessels in vivo. Nat Biotechnol, 2007. 25(3): p. 317–8.PubMedCrossRefGoogle Scholar
  11. 11.
    Yamahara, K., et al., Augmentation of neovascularization [corrected] in hindlimb ischemia by combined transplantation of human embryonic stem cells-derived endothelial and mural cells. PLoS One, 2008. 3(2): p. e1666.PubMedCrossRefGoogle Scholar
  12. 12.
    Levenberg, S., et al., Endothelial cells derived from human embryonic stem cells. Proc Natl Acad Sci USA, 2002. 99(7): p. 4391–6.PubMedCrossRefGoogle Scholar
  13. 13.
    Chen, T., et al., Stromal cell-derived factor-1/CXCR4 signaling modifies the capillary-like organization of human embryonic stem cell-derived endothelium in vitro. Stem Cells, 2007. 25(2): p. 392–401.PubMedCrossRefGoogle Scholar
  14. 14.
    Li, Z., et al., Comparison of reporter gene and iron particle labeling for tracking fate of human embryonic stem cells and differentiated endothelial cells in living subjects. Stem Cells, 2008. 26(4): p. 864–73.PubMedCrossRefGoogle Scholar
  15. 15.
    Feraud, O., Y. Cao, and D. Vittet, Embryonic stem cell-derived embryoid bodies development in collagen gels recapitulates sprouting angiogenesis. Lab Invest, 2001. 81(12): p. 1669–81.PubMedGoogle Scholar
  16. 16.
    Li, Z., et al., Functional and transcriptional characterization of human embryonic stem cell-derived endothelial cells for treatment of myocardial infarction. PLoS One, 2009. 4(12): p. e8443.PubMedCrossRefGoogle Scholar
  17. 17.
    Li, Z., et al., Functional characterization and expression profiling of human induced ­pluripotent stem cell- and embryonic stem cell-derived endothelial cells. Stem Cells Dev, 2011. [Epub ahead of print]PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press 2011

Authors and Affiliations

  • Shijun Hu
    • 1
    • 2
  • Preston Lavinghousez
    • 1
    • 2
  • Zongjin Li
    • 1
    • 2
  • Joseph C. Wu
    • 1
    • 2
    • 3
    Email author
  1. 1.Department of Medicine, Division of CardiologyStanford University School of MedicineStanfordUSA
  2. 2.Department of RadiologyStanford University School of MedicineStanfordUSA
  3. 3.Institute of Stem Cell Biology and Regenerative MedicineStanford University School of MedicineStanfordUSA

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