Abstract
Endothelial cells (ECs), as important constituent of blood vessel are crucial to maintaining vascular tone, and any dysfunction in ECs could lead to many diseases of the vascular system. Due to limited availability of human samples, most of our research has been limited to animal models. Induced pluripotent stem cells (iPSCs) due to their potential to differentiate into any cell type of the human body, including cells of the cardiovascular system, provide us with a great opportunity to study human vascular disease. This protocol describes an efficient, simple, and step-wise method to differentiate iPSCs to endothelial cells (iPSC-ECs) using small molecules and growth factors. Through the activation of WNT signaling pathway, iPSCs are differentiated towards a mesodermal lineage with the potential to generate endothelial progenitors. These progenitors are further stimulated with growth factors to generate endothelial cells (Figure 1A).Figure 1. Stepwise differentiation strategy for iPSCs endothelial cell differentiation. A. chemicals/growth factors used at each stage of differentiation. B. Morphology of cells at different stage of differentiation. Scale bars = 500 µm.
Keywords: iPSCs, Endothelial cells, Differentiation
Materials and Reagents
Equipment
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Acknowledgments
The work was supported through funds from by the American Heart Association Postdoctoral Grant (16POST30960020) to Chun Liu; the National Institutes of Health (K01 HL135455), Stanford Translational Research and Applied Medicine scholar grant to Nazish Sayed. This protocol is based on our previous work (Matsa et al., 2016).
Competing interests
Authors have no conflict of interest or competing interest.
References
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or 2%?
10 ml B27 minus insulin into 500 ml RPMI medium, 2%
Thanks!
Hi Laura,We add more VEGF and bFGF in addition to the EGM-2 supplement.
Ok great, thank you very much for your reply!