Nguyen Thi Minh Xuan, Vegrichtova Marketa, Tlapakova Tereza, Krulova Magdalena, Krylov Vladimir
Department of Cell Biology, Charles University, Faculty of Science, Vinicna 7, Prague 2 128 44, Czech Republic.
Stem Cells Int. 2019 May 5;2019:8387478. doi: 10.1155/2019/8387478. eCollection 2019.
Epithelial-mesenchymal transition (EMT) is a fundamental process in embryonic development by which sessile epithelial cells are converted into migratory mesenchymal cells. Our laboratory has been successful in the establishment of immature Sertoli cells (XtiSCs) with the restricted differentiation potential. The aim of this study is the determination of factors responsible for EMT activation in XtiSCs and stemness window acquisition where cells possess the broadest differentiation potential. For this purpose, we tested three potent EMT inducers-GSK-3 inhibitor (CHIR99021), FGF2, and/or TGF-1 ligand. XtiSCs underwent full EMT after 3-day treatment with CHIR99021 and partial EMT with FGF2 but not with TGF-1. The morphological change of CHIR-treated XtiSCs to the typical spindle-like cell shape was associated with the upregulation of mesenchymal markers and the downregulation of epithelial markers. Moreover, only CHIR-treated XtiSCs were able to differentiate into chondrocytes and cardiomyocytes . Interestingly, EMT-shifted cells could migrate towards cancer cells (HeLa) and to the injury site . The results provide a better understanding of signaling pathways underlying the generation of testis-derived stem cells.
上皮-间质转化（EMT）是胚胎发育过程中的一个基本过程，通过该过程，固着的上皮细胞转化为迁移性间充质细胞。我们实验室已成功建立了具有有限分化潜能的未成熟支持细胞（XtiSCs）。本研究的目的是确定在XtiSCs中激活EMT以及细胞具有最广泛分化潜能时获得干性窗口的相关因素。为此，我们测试了三种有效的EMT诱导剂——GSK-3抑制剂（CHIR99021）、FGF2和/或TGF-1配体。用CHIR99021处理3天后，XtiSCs发生了完全EMT，用FGF2处理则发生了部分EMT，而用TGF-1处理则未发生EMT。CHIR处理的XtiSCs形态转变为典型的纺锤状细胞形状，这与间充质标志物的上调和上皮标志物的下调有关。此外，只有CHIR处理的XtiSCs能够分化为软骨细胞和心肌细胞。有趣的是，发生EMT转变的细胞能够向癌细胞（HeLa）和损伤部位迁移。这些结果有助于更好地理解睾丸来源干细胞生成背后的信号通路。
