文献类型：期刊文献

英文题名：Differentiation roadmap of embryonic Sertoli cells derived from mouse embryonic stem cells

作者：Xu, Chenze[1];Mohsin, Ali[1];Luo, Yanxia[1];Xie, Lili[1];Peng, Yan[1];Wang, Qizheng[1];Hang, Haifeng[1,2];Zhuang, Yingping[1,2];Guo, Meijin[1,2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Minist Educ, Engn Res Ctr Proc Syst, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2019

卷号：10

外文期刊名：STEM CELL RESEARCH & THERAPY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000460812000003)】；

基金：This research was supported from the National Natural Science Foundation of China (81373286) and was also funded by the Fundamental Research Funds for the China Central Universities (No. 22221818014 and No. 22221817014).

语种：英文

外文关键词：Embryonic stem cells; Embryonic Sertoli cells; Lentiviral transduction; Molecular mechanism; Gonadogenesis; Male determinant factors

摘要：BackgroundEmbryonic Sertoli cells (eSCs) play an important role in sex determination and in male gonad development which makes them a very useful cell type for therapeutic applications. However, the deriving mechanism of Sertoli cells has been unclear and challenging to create a large number of quality eSCs. Therefore, this study aimed to create the eSCs induced from mouse embryonic stem (mES) cells by regulating defined factors and to explore the relevant regulatory mechanism.MethodsSix inducing factors, Sry, Sox9, SF1, WT1, GATA4, and Dmrt1, were respectively transduced into mES cells by lentiviral infection according to the experimental design. The test groups were identified by development stage-specific markers, AMH, Emx2, SF1, and FasL, using flow cytometry. Induced eSCs were determined by FasL and AMH biomarkers under immunofluorescence, immunocytochemistry, and flow cytometry. Moreover, the pluripotency markers, gonad development-related markers, epithelialmarkers and mesenchymal markers in test groups were transcriptionally determined by qPCR.ResultsIn this study, the co-overexpression of all the six factors effectively produced a large population of eSCs from mES cells in 35days of culturing. These eSCs were capable of forming tubular-like and ring-like structures with functional performance. The results of flow cytometry indicated that the upregulation of GATA4 and WT1 contributed to the growth of somatic cells in the coelomic epithelium regarded as the main progenitor cells of eSCs. Whereas,SF1 facilitated the development of eSC precursor cells, and Sry and Sox9 promoted the determination of male development. Moreover, the overexpression of Dmrt1 was essential for the maintenance of eSCs and some of their specific surface biomarkers such as FasL. The cellular morphology, biomarker identification, and transcriptomic analysis aided in exploring the regulatory mechanism of deriving eSCs from mES cells.ConclusionConclusively, we have elucidated a differentiation roadmap of eSCs derived from mEScells with a relevant regulatory mechanism. Through co-overexpression of all these six factors, a large population of eSCs was successfully induced occupying 24% of the whole cell population (1x10(5) cells/cm(2)). By adopting this approach, a mass of embryonic Sertoli cells can be generated for the purpose of co-culture technique, organ transplantation, gonadal developmental and sex determination researches.