ETV2 Overexpression Promotes Efficient Differentiation of Pluripotent Stem Cells to Endothelial Cells

IF 3.5 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology and Bioengineering Pub Date : 2025-03-25 DOI:10.1002/bit.28979

Yunfeng Ding, Soniya Tamhankar, Feifan Du, Tessa Christopherson, Nate Schlueter, Jenna R. Cohen, Eric V. Shusta, Sean P. Palecek

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引用次数: 0

Abstract

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.

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从人多能干细胞（hPSCs）分化出内皮细胞（ECs）通常需要两周时间，而且需要优化参数。在hPSCs中过表达细胞类型特异性转录因子可高效分化成各种细胞类型。ETV2是决定内皮命运的关键转录因子，在hPSCs中过表达可诱导快速、简便的EC分化（iETV2-ECs）。我们开发了一种两阶段策略，即第一阶段在基础诱导培养基中分化诱导性 ETV2-过表达 hPSCs，第二阶段在内皮培养基中扩增它们。通过优化播种密度和培养基成分，我们在5天内就获得了99%纯度的CD31+ CD144+ iETV2-ECs，而无需进行细胞分拣。iETV2-ECs表现出体外血管生成潜能、低密度脂蛋白摄取和细胞因子反应。转录组比较显示，iETV2-ECs 和传统分化 ECs 的基因表达谱相似。此外，iETV2-ECs 对 Wnt 信号激动剂和 TGFβ 抑制剂有反应，获得了脑 EC 表型，使其成为血脑屏障建模等应用的可扩展 EC 来源。

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来源期刊

Biotechnology and Bioengineering 工程技术-生物工程与应用微生物

CiteScore

7.90

自引率

5.30%

发文量

280

审稿时长

2.1 months

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