ETV2 overexpression promotes efficient differentiation of pluripotent stem cells to endothelial cells

ABSTRACT: Directed differentiation of endothelial cells (ECs) from human pluripotent stem cells (hPSCs) typically takes around fourteen days, requires optimization of parameters such as hPSC seeding density and medium supplement concentration or timing, and often necessitates cell sorting to achieve high purity ECs. Overexpression of cell type-specific transcription factors in hPSCs has been shown to lead to efficient differentiation of many cell types, including neurons, astrocytes and hepatocytes. ETV2 has been identified as an essential transcription factor for endothelial cell fate commitment during development. Here, we present a simple, well-characterized differentiation method to generate ECs by overexpressing ETV2 in an inducible manner (iETV2) in hPSC cell lines. The result is a two-stage differentiation strategy, in which ECs are differentiated in a basal medium during stage I and then expanded in endothelial medium during stage II. By optimizing hPSC seeding density and differentiation medium, we generated an EC population with more than 99% pure CD31+ CD144+ cells without any cell sorting in just five days. In vitro angiogenesis potential of the iETV2-ECs was confirmed in cord formation assays and iETV2-EC uptake of acetylated LDL and response to inflammatory cytokines was confirmed. We compared transcriptomics of iETV2-ECs to the ECs generated from hPSCs via directed differentiation and confirmed similarity of gene expression profiles of ECs generated by these two different approaches. Furthermore, we found that iETV2-ECs respond to previously identified Wnt signaling agonist CHIR99021 and TGFβ signaling inhibitor RepSox to acquire aspects of brain EC phenotype. These findings underscore that iETV2-ECs can serve as easily scalable endothelial source, powering a variety of research applications, potentially including in vitro blood-brain barrier modeling.

ORGANISM(S): Homo sapiens

PROVIDER: GSE271101 | GEO | 2025/03/24

REPOSITORIES: GEO