Induction of Sertoli-like cells from human fibroblasts by NR5A1 and GATA4.

Jianlin Liang,Wenbo Wu,Jing He,Jian Du,Kehkooi Kee,Yahui Guo,Nan Wang,Chencheng Yao,Lin Li,Zheng Li

doi:10.7554/elife.48767

Abstract

Sertoli cells are essential nurse cells in the testis that regulate the process of spermatogenesis and establish the immune-privileged environment of the blood-testis-barrier (BTB). Here, we report the in vitro reprogramming of fibroblasts to human induced Sertoli-like cells (hiSCs). Initially, five transcriptional factors and a gene reporter carrying the AMH promoter were utilized to obtain the hiSCs. We further reduce the number of reprogramming factors to two, NR5A1 and GATA4, and show that these hiSCs have transcriptome profiles and cellular properties that are similar to those of primary human Sertoli cells. Moreover, hiSCs can sustain the viability of spermatogonia cells harvested from mouse seminiferous tubules. hiSCs suppress the proliferation of human T lymphocytes and protect xenotransplanted human cells in mice with normal immune systems. hiSCs also allow us to determine a gene associated with Sertoli cell only syndrome (SCO), CX43, is indeed important in regulating the maturation of Sertoli cells.

Highlights

Sertoli cells are the first somatic cell type to differentiate in the testis and the only somatic cell type inside the seminiferous tubules
Testing the reprogramming capability of five putative transcriptional factors Based on the reprogramming capability of the transcriptional factors reported in a mouse study (Buganim et al, 2012), we first tested the reprogramming capabilities of the human homologs of the five transcriptional factors (5TFs: NR5A1, GATA4, WT1, SOX9 and DMRT1) to convert human fibroblasts to human induced Sertoli-like cells (hiSCs)
To further confirm that the AMH:EGFP + 5F-hiSCs have the signature of Sertoli cells, we examined the expression of several Sertoli cell markers, including CLU, NCAM2, DHH, ERBB4, INHB, INHA, SHBG, GATA6, CDKN1B, TGFa and LMMA3, by quantitative PCR

Summary

Introduction

Sertoli cells are the first somatic cell type to differentiate in the testis and the only somatic cell type inside the seminiferous tubules. Sertoli cells play a critical role in directing testis morphogenesis and the creation of an immune-privileged microenvironment, which is required for male germ cell development. NR5A1 (or SF1), GATA4, and WT1 are major transcriptional factors that direct somatic cells to become fetal Sertoli cells (Rotgers et al, 2018). Five transcriptional factors have been demonstrated to successfully reprogram mouse fibroblasts to Sertoli cells (Buganim et al, 2012). The expanding fetal Sertoli cells and another type of testicular somatic cell (i.e., peritubular cells) regulate the final organization and morphogenesis of the developing gonad into a testis (Griswold, 1998; McLaren, 2000)

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