Endothelial Progenitor Cells Produced From Human Pluripotent Stem Cells by a Synergistic Combination of Cytokines, Small Compounds, and Serum-Free Medium

Frontiers in Cell and Developmental Biology
Simon FarkasIrena Koutna

Abstract

Human pluripotent stem cells (hPSCs) are a promising source of autologous endothelial progenitor cells (EPCs) that can be used for the treatment of vascular diseases. However, this kind of treatment requires a large amount of EPCs. Therefore, a highly efficient, robust, and easily reproducible differentiation protocol is necessary. We present a novel serum-free differentiation protocol that exploits the synergy of multiple powerful differentiation effectors. Our protocol follows the proper physiological pathway by differentiating EPCs from hPSCs in three phases that mimic in vivo embryonic vascular development. Specifically, hPSCs are differentiated into (i) primitive streak, which is subsequently turned into (ii) mesoderm, which finally differentiates into (iii) EPCs. This differentiation process yields up to 15 differentiated cells per seeded hPSC in 5 days. Endothelial progenitor cells constitute up to 97% of these derived cells. The experiments were performed on the human embryonic stem cell line H9 and six human induced pluripotent stem cell lines generated in our laboratory. Therefore, robustness was verified using many hPSC lines. Two previously established protocols were also adapted and compared to our synergistic thre...Continue Reading

References

Mar 31, 2000·Nature Medicine·P Carmeliet
Nov 30, 2010·Cell Stem Cell·Maxim A VodyanikIgor I Slukvin
Sep 21, 2013·Stem Cell Reports·William J AdamsGuillermo García-Cardeña
Jul 28, 2015·Nature Cell Biology·Christoph PatschChad A Cowan
Mar 10, 2016·Stem Cells Translational Medicine·Mark Seow Khoon ChongJerry Kok Yen Chan

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Citations

Aug 14, 2020·Bioengineering·Mariana A BrancoMaria Margarida Diogo
Jul 23, 2021·American Journal of Respiratory Cell and Molecular Biology·Olena A KolesnichenkoVladimir V Kalinichenko

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Methods Mentioned

BETA
by light microscopy
florescence-activated cell sorting
FACS
flow cytometry
light microscopy
PCR

Software Mentioned

Flowing
BD FACSDiva

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