Inducing Non-genetically Modified Induced Embryonic Sertoli Cells Derived From Embryonic Stem Cells With Recombinant Protein Factors

Chenze Xu; Ali Mohsin; Yanxia Luo; Lili Xie; Yan Peng; Qizheng Wang; Waqas Ahmed; Haifeng Hang; Yingping Zhuang; Meijin Guo

doi:10.3389/fcell.2020.533543

Inducing Non-genetically Modified Induced Embryonic Sertoli Cells Derived From Embryonic Stem Cells With Recombinant Protein Factors

Front Cell Dev Biol. 2021 Jan 25:8:533543. doi: 10.3389/fcell.2020.533543. eCollection 2020.

Authors

Chenze Xu¹, Ali Mohsin¹, Yanxia Luo¹, Lili Xie¹, Yan Peng¹, Qizheng Wang¹, Waqas Ahmed¹, Haifeng Hang^{1

2}, Yingping Zhuang^{1

2}, Meijin Guo^{1

2

3}

Affiliations

¹ State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
² Engineering Research Centre of Processes System, Ministry of Education, East China University of Science and Technology, Shanghai, China.
³ Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China.

Abstract

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 Sry, Sox9, Sf1, Wt1, Gata4, and Dmrt1. 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.

Keywords: Sertoli cells; embryonic stem cells; lentiviral transduction; non-genetically modified; recombinant endogenous protein factors.