Protocol

Progenitor Cells

Volume 916 of the series Methods in Molecular Biology pp 81-96

Date:

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

  • S. Bahram BahramiAffiliated withDepartment of Surgery, University of California San Francisco
  • , Mandana VeisehAffiliated withLife Science Division, Lawrence Berkeley National Laboratory
  • , Nancy J. BoudreauAffiliated withDepartment of Surgery, University of California San Francisco Email author 

* Final gross prices may vary according to local VAT.

Get Access

Abstract

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