Mapping molecular pathways for embryonic Sertoli cells derivation based on differentiation model of mouse embryonic stem cells

Chenze Xu,Ali Mohsin,Yichen Dai,Haifeng Hang,Yingping Zhuang,Meijin Guo

doi:10.1186/s13287-020-01600-2

Abstract

BackgroundEmbryonic Sertoli cells (eSCs) have been known for playing important roles in male reproductive development system. In current studies, eSCs were mainly generated from induced intermediate mesoderm. The deriving mechanism of eSCs has been unclear so far. Therefore, this work was aimed to reveal the molecular pathways during derivation of eSCs.MethodsIn this scenario, a differentiation model from mouse embryonic stem cells (mESCs) to eSCs was established through spatiotemporal control of 5 key factors, Wilms tumor 1 homolog (Wt1), GATA binding protein 4 (Gata4), nuclear receptor subfamily 5, group A, member 1 (Nr5a1, i.e., Sf1), SRY (sex determining region Y)-box 9 (Sox9), doublesex, and mab-3 related transcription factor 1 (Dmrt1). To investigate the molecular mechanism, these key factors were respectively manipulated through a light-switchable (light-on) system, tetracycline-switchable (Tet-on) system, and CRISPR/Cas9 knock out (KO) system.ResultsVia the established approach, some embryonic Sertoli-like cells (eSLCs) were induced from mESCs and formed ring-like or tubular-like structures. The key factors were respectively manipulated and revealed their roles in the derivation of these eSLCs. Based on these results, some molecular pathways were mapped during the development of coelomic epithelial somatic cells to eSCs.ConclusionsThis differentiation model provided a high controllability of some key factors and brought a novel insight into the deriving mechanism of Sertoli cells.

Highlights

Embryonic Sertoli cells have been known for playing important roles in male reproductive development system
SGPCs were the common precursor cells of male gonad supporting cells (Sertoli cells) and female gonadal supporting cells [6, 16]. These SGPCs developed into embryonic Sertoli cell under the influence of factors including Wilms tumor 1 homolog (Wt1), GATA binding protein 4 (Gata4), Sry, SRY-box 9 (Sox9), SRY-box 8 (Sox8), fibroblast growth factor 9 (Fgf9), fibroblast growth factor receptor 2 (Fgfr2), prostaglandin D2 (PGD2), prostaglandin D2 synthase (Ptgds), antiMullerian hormone (Amh), doublesex and mab-3 related transcription factor 1 (Dmrt1), and glial cell line derived neurotrophic factor (Gdnf) [16,17,18,19,20,21]
Determination of established differentiation model from mouse embryonic stem cells (mESCs) to Embryonic Sertoli cells (eSCs) In order to map the molecular differentiation pathways for deriving eSCs, this work aimed to establish a differentiation from mESCs to eSCs to reproduce the expression schedule of key factors and analyze cellular morphology and organic forms

Summary

Introduction

Embryonic Sertoli cells (eSCs) have been known for playing important roles in male reproductive development system. Xu et al Stem Cell Research & Therapy (2020) 11:85 protein kinase 4 (Map3k4), growth arrest, and DNAdamage-inducible 45 gamma (Gadd45g) [6,7,8,9,10,11,12,13,14,15] In mice, these SF1-positive precursor cells (SPCs) ingressed into bipotential gonads and became SF1-positive gonadal precursor cells (SGPCs, i.e., pre-Sertoli cells) in 11.2–11.4 day post coitum (dpc). Many theories on the deriving mechanism of Sertoli cells have already been proposed; still further investigation is needed to explore the complete mechanism of deriving eSCs [1, 6, 16, 22]

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