The ability to differentiate human pluripotent stem cells into endothelial cells with properties of cord-blood endothelial colony–forming cells (CB-ECFCs) may enable the derivation of clinically relevant numbers of highly proliferative blood vessel–forming cells to restore endothelial function in patients with vascular disease. We describe a protocol to convert human induced pluripotent stem cells (hiPSCs) or embryonic stem cells (hESCs) into cells similar to CB-ECFCs at an efficiency of >108 ECFCs produced from each starting pluripotent stem cell. The CB-ECFC-like cells display a stable endothelial phenotype with high clonal proliferative potential and the capacity to form human vessels in mice and to repair the ischemic mouse retina and limb, and they lack teratoma formation potential. We identify Neuropilin-1 (NRP-1)-mediated activation of KDR signaling through VEGF165 as a critical mechanism for the emergence and maintenance of CB-ECFC-like cells.
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These studies were supported by funds provided by the Riley Children's Foundation (M.C.Y.), by an AHA postdoctoral fellowship (N.P.), and Public Health Service Grants R01 HL109602 (M.C.Y. and S.L.V.-H.), RO1 HL056416 (H.E.B.), RO1 HL067384 (H.E.B.), PO1 DK090948 (H.E.B. and M.C.Y.), and by the Bio & Medical Technology Development Program of the National Research Foundation (NRF), which is funded by the South Korean government (MEST; No. 2011-0019487). The authors thank H.P. Poudel (Indiana University Purdue University Indianapolis) for critical input in biostatistical analysis. The authors also thank H.J. Lee (CHA Stem Cell Institute, CHA University, South Korea) for HLI experiments.
The authors declare no competing financial interests.
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Prasain, N., Lee, M., Vemula, S. et al. Differentiation of human pluripotent stem cells to cells similar to cord-blood endothelial colony–forming cells. Nat Biotechnol 32, 1151–1157 (2014). https://doi.org/10.1038/nbt.3048
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