Mouse embryonic stem cells (mESCs) can maintain self-renewal and differentiate into any cell type of the three primary germ layers. The vascular endothelial growth factor (VEGF) is involved in the regulation of mESC differentiation and induces the activation of a series of kinase responses and several cell signaling pathways by binding to its respective transmembrane receptors, vascular endothelial growth factor receptor VEGFR1, and VEGFR2. Fruquintinib is a selective inhibitor of VEGFRs, and we used it to investigate the effects on the maintenance of pluripotency and differentiation potential of mESCs in this study. Our results showed that fruquintinib-treated cells expressed higher levels of pluripotent markers, including Oct4, Nanog, Sox2, and Esrrb under serum and leukemia inhibitory factor (LIF) condition, whereas the expression of phosphorylated Erk1/2 was restricted. Mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (MEK) signaling inhibitor (PD0325901) and glycogen synthase kinase 3 (GSK3) signaling inhibitor (CHIR99021) (also known as 2i) enable cells to maintain naive pluripotency with LIF, and fruquintinib can also promote cells to maintain naive pluripotent state even under serum/LIF condition, whereas VEGF addition limits the pluripotency characteristics in serum/LIF mESCs. Furthermore, fruquintinib could inhibit the three-germ layer establishment in embryoid body formation and maintain the undifferentiated characteristics of mESCs, indicating that fruquintinib could promote the maintenance of naive pluripotency and inhibit early differentiation programs.

中文翻译：

---

呋喹替尼对小鼠胚胎干细胞多能性维持和分化潜能的影响

小鼠胚胎干细胞 (mESCs) 可以维持自我更新并分化成三个主要胚层的任何细胞类型。血管内皮生长因子 (VEGF) 参与调节 mESC 分化并通过与其各自的跨膜受体、血管内皮生长因子受体 VEGFR1 和 VEGFR2 结合来诱导一系列激酶反应和几种细胞信号通路的激活。呋喹替尼是一种选择性 VEGFRs 抑制剂，我们在本研究中用它来研究对维持 mESCs 多能性和分化潜能的影响。我们的研究结果表明，在血清和白血病抑制因子 (LIF) 条件下，呋喹替尼处理的细胞表达更高水平的多能标志物，包括 Oct4、Nanog、Sox2 和 Esrrb，而磷酸化 Erk1/2 的表达受到限制。丝裂原活化蛋白激酶 (MAPK)/细胞外信号调节激酶 (MEK) 信号抑制剂 (PD0325901) 和糖原合酶激酶 3 (GSK3) 信号抑制剂 (CHIR99021)（也称为 2i）使细胞能够通过 LIF 维持幼稚多能性，即使在血清/LIF 条件下，呋喹替尼也可以促进细胞维持幼稚多能状态，而 VEGF 的添加限制了血清/LIF mESCs 的多能性特征。此外，呋喹替尼可抑制胚状体形成过程中三胚层的建立，维持mESCs的未分化特性，表明呋喹替尼可促进幼稚多能性的维持，抑制早期分化程序。丝裂原活化蛋白激酶 (MAPK)/细胞外信号调节激酶 (MEK) 信号抑制剂 (PD0325901) 和糖原合酶激酶 3 (GSK3) 信号抑制剂 (CHIR99021)（也称为 2i）使细胞能够通过 LIF 维持幼稚多能性，即使在血清/LIF 条件下，呋喹替尼也可以促进细胞维持幼稚多能状态，而 VEGF 的添加限制了血清/LIF mESCs 的多能性特征。此外，呋喹替尼可抑制胚状体形成过程中三胚层的建立，维持mESCs的未分化特性，表明呋喹替尼可促进幼稚多能性的维持，抑制早期分化程序。丝裂原活化蛋白激酶 (MAPK)/细胞外信号调节激酶 (MEK) 信号抑制剂 (PD0325901) 和糖原合酶激酶 3 (GSK3) 信号抑制剂 (CHIR99021)（也称为 2i）使细胞能够通过 LIF 维持幼稚多能性，即使在血清/LIF 条件下，呋喹替尼也可以促进细胞维持幼稚多能状态，而 VEGF 的添加限制了血清/LIF mESCs 的多能性特征。此外，呋喹替尼可抑制胚状体形成过程中三胚层的建立，维持mESCs的未分化特性，表明呋喹替尼可促进幼稚多能性的维持，抑制早期分化程序。