ETV2 Overexpression Promotes Efficient Differentiation of Pluripotent Stem Cells to Endothelial Cells
Summary
Differentiating endothelial cells (ECs) from human pluripotent stem cells (hPSCs) typically takes 2 weeks and requires parameter optimization. Overexpression of cell type-specific transcription factors in hPSCs has shown efficient differentiation into various cell types. ETV2, a crucial transcription factor for endothelial fate, can be overexpressed in hPSCs to induce rapid and facile EC differentiation (iETV2-ECs). We developed a two-stage strategy which involves differentiating inducible ETV2-overexpressing hPSCs in a basal induction medium during stage I and expanding them in an endothelial medium during stage II. By optimizing seeding density and medium composition, we achieved 99% pure CD31+ CD144+ iETV2-ECs without cell sorting in 5 days. iETV2-ECs demonstrated in vitro angiogenesis potential, LDL uptake, and cytokine response. Transcriptomic comparisons revealed similar gene expression profiles between iETV2-ECs and traditionally differentiated ECs. Additionally, iETV2-ECs responded to Wnt signaling agonist and TGFβ inhibitor to acquire brain EC phenotypes, making them a scalable EC source for applications including blood-brain barrier modeling. © 2025 The Author(s). Biotechnology and Bioengineering published by Wiley Periodicals LLC.
| Authors | Ding Y, Tamhankar S, Du F, Christopherson T, Schlueter N, Cohen JR, Shusta EV, Palecek SP |
|---|---|
| Journal | Biotechnology and bioengineering |
| Publication Date | 2025 Jul;122(7):1914-1928 |
| PubMed | 40134129 |
| PubMed Central | PMC12152468 |
| DOI | 10.1002/bit.28979 |