Isolation and Expansion of Endothelial Progenitor Cells Derived from Mouse Embryonic Stem Cells

  • S. Bahram Bahrami
  • Mandana Veiseh
  • Nancy J. Boudreau
Part of the Methods in Molecular Biology book series (MIMB, volume 916)


The unlimited differentiation and proliferation capacity of embryonic stem cells represents a great resource for regenerative medicine. Here, we describe a method for differentiating, isolating, and expanding endothelial cells (ECs) from mouse embryonic stem cells (mESCs). First, mESCs are expanded on a mouse embryonic fibroblast (mEF) feeder layer and partially differentiated into embryoid bodies (EBs) by growing the cells in an ultra-low attachment plate for up to 5 days. The EBs are then differentiated along the endothelial lineage using endothelial growth medium supplemented with 40 ng/mL vascular endothelial growth factor (VEGF). The differentiated endothelial population expresses both Fetal Liver Kinase 1 (Flk-1) and VE-Cadherin on the cell surface which can be further purified using a fluorescence-activated cell sorting (FACS) system and subsequently expanded on 0.1 % gelatin-coated plates. The differentiated cells can be analyzed by real-time PCR and flow cytometry to confirm enrichment of EC-specific genes and proteins.

Key words

Embryonic stem cells Differentiation FACS Endothelial cells 



This work is supported by grants from NIH/NCI TMEN grant (U54CA126552.) to Nancy Boudreau and Mina J Bissell and U.S. Department of Energy, Office of Biological and Environmental Research (DE-AC02-05CH1123), a Distinguished Fellow Award and Low Dose Radiation Program (03-76SF00098) to Mina J. Bissell. Mandana Veiseh was supported by a postdoctoral fellowship from the NCI of the NIH (F32 CA132491A). We thank Pamela Derish in the Department of Surgery at UCSF for editorial review of the manuscript.


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Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • S. Bahram Bahrami
    • 1
  • Mandana Veiseh
    • 2
  • Nancy J. Boudreau
    • 1
  1. 1.Department of SurgeryUniversity of California San FranciscoSan FranciscoUSA
  2. 2.Life Science Division, Lawrence Berkeley National LaboratoryBerkeleyUSA

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