| Chapter title |
Isolation and expansion of endothelial progenitor cells derived from mouse embryonic stem cells.
|
|---|---|
| Chapter number | 7 |
| Book title |
Progenitor Cells
|
| Published in |
Methods in molecular biology, January 2012
|
| DOI | 10.1007/978-1-61779-980-8_7 |
| Pubmed ID | |
| Book ISBNs |
978-1-61779-979-2, 978-1-61779-980-8
|
| Authors |
Bahrami SB, Veiseh M, Boudreau NJ, S. Bahram Bahrami, Mandana Veiseh, Nancy J. Boudreau |
| 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. |
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