Xu Chenze, Mohsin Ali, Luo Yanxia, Xie Lili, Peng Yan, Wang Qizheng, Ahmed Waqas, Hang Haifeng, Zhuang Yingping, Guo Meijin
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
Engineering Research Centre of Processes System, Ministry of Education, East China University of Science and Technology, Shanghai, China.
Front Cell Dev Biol. 2021 Jan 25;8:533543. doi: 10.3389/fcell.2020.533543. eCollection 2020.
Embryonic Sertoli cells (eSCs) possess multiple supporting functions and research value in gonadal development and sex determination. However, the limitation of acquiring quality eSCs had hindered the further application. Herein, we successfully derived non-genetically modified (non-GM)-induced embryonic Sertoli-like cells (eSLCs) from mouse embryonic stem cells (ESCs) with a TM4 cell-derived conditioned medium containing recombinant endogenous protein factors , , , , , and . These eSLCs were determined through morphology; transcriptional expression levels of stage-specific, epithelial, and mesenchymal marker genes; flow cytometry, immunofluorescence; and immunocytochemistry and functionally determined by coculture with spermatogonia stem cells. Results indicated that these eSLCs performed similarly to eSCs in specific biomarkers and expression of marker genes and supported the maturation of spermatogonia. The study induced eSLCs from mouse ESCs by defined protein factors. However, the inducing efficiency of the non-GM method was still lower than that of the lentiviral transduction method. Thus, this work established a foundation for future production of non-GM eSLCs for clinical applications and fundamental theory research.
胚胎支持细胞（eSCs）在性腺发育和性别决定中具有多种支持功能和研究价值。然而，获取高质量eSCs的局限性阻碍了其进一步应用。在此，我们利用含有重组内源性蛋白因子、、、、、和的TM4细胞条件培养基，成功从小鼠胚胎干细胞（ESCs）中诱导出非转基因（non-GM）的胚胎支持样细胞（eSLCs）。这些eSLCs通过形态学、阶段特异性、上皮和间充质标记基因的转录表达水平、流式细胞术、免疫荧光以及免疫细胞化学进行鉴定，并通过与精原干细胞共培养进行功能鉴定。结果表明，这些eSLCs在特定生物标志物和标记基因表达方面与eSCs表现相似，并支持精原细胞成熟。该研究通过特定蛋白因子从小鼠ESCs中诱导出eSLCs。然而，非转基因方法的诱导效率仍低于慢病毒转导方法。因此，这项工作为未来生产用于临床应用和基础理论研究的非转基因eSLCs奠定了基础。
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