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Methods Mol Biol. 2012;916:81-96. doi: 10.1007/978-1-61779-980-8_7.

Isolation and expansion of endothelial progenitor cells derived from mouse embryonic stem cells.

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  • 1Department of Surgery, University of California San Francisco, San Francisco, CA, USA.

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.

PMID:
22914934
[PubMed - indexed for MEDLINE]
PMCID:
PMC3733326
Free PMC Article

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