Differentiation of Murine Embryonic Stem Cells into Endothelial Cells

Objective: In this investigation Murine Embryonic Stem (ES) cells were differentiated into endothelial cells. Materials and Methods: Murine ES cells (CCE cell line) exposed to AlphaMEM medium containing 10% FBS for 4 days. Then obtained Flk-1 (Flk-1: Vascular Endothelial Growth Factor Receptor 2) Positive cells were cultuted in Endothelial Growth Medium-2 (EGM-2) until the last day of experiment. Differentiated cells were evaluated by immunocytochemistry, RT-PCR and Tube Formation Assays. Results: When the ES cells cultured in collagen coated dishes containing Alpha-MEM & FBS, Flk-1 positive cells were obtained. After transfering Flk-1 positive cells into fibronectin coated dishes containing EGM2, the cells were assumed a relatively uniform endothelial cell morphology and could be propagated and expanded. Immunocytochemical and RT-PCR analysis of differentiated cells showed that they take up acetylated low-density lipoprotein (LDL), express Flk-1, CD31 and bind the BS-l lectin. When placed in Matrigel, these Murine ES cell–derived endothelial cells formed capillary-like structures characteristic of endothelial cells Conclusion: ES cell–derived endothelial cells provide a novel means to examine the mechanisms of endothelial cell development, and may open up new therapeutic strategies.