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

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

doi:10.3389/fcell.2020.533543

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.

Highlights

Sertoli cells, as the main supporting cells of testes, have important roles in male determination, male gonadal development, and normal testicular functioning (Skinner and Griswold, 2005; Carre and Greenfield, 2016; Gunes et al, 2016)
These limitations hamper their application of large-scale production of embryonic stem cells (ESCs)
We proposed to add the concentrated supernatant of TM4 cells for improving the ESC culture

Summary

Introduction

As the main supporting cells of testes, have important roles in male determination, male gonadal development, and normal testicular functioning (Skinner and Griswold, 2005; Carre and Greenfield, 2016; Gunes et al, 2016). They possess high clinical importance due to their immunosuppression function for transplantation application (Luca et al, 2018). ESCs are of great value for fundamental theory research and future clinical application

Methods

Results

Discussion

Conclusion