In Vitro Derivation of Functional Sertoli-Like Cells from Mouse Embryonic Stem Cells

Abstract

Sertoli cells (SCs) in the mammalian testes are well known as supporting cells of spermatogenesis, but have recently become an attractive source of cell therapy because of their capacity for immune modulation and trophic effects. In order to increase their applicable efficacy, we demonstrate a novel differentiation method for mouse embryonic stem cell (ESC)-derived Sertoli-like cells (SLCs) via the intermediate mesoderm (IM). We show that IM derived from an induction of 6 days expressed markers such as Wt1, Lhx1, Pax2 and Osr1, and that a sequential induction of 6 days resulted in ESC-SLCs. The SLCs expressed their marker genes (Sf1, Sox9, Gata4, Wt1, Fshr and Scf), but the pluripotency-marker gene Oct4 was decreased. After sorting by FSHR expression, high-purity (> 90%) SLCs were collected that showed distinct characteristics of SCs such as high phagocytic and immune modulation activities as well as the expression of immune-related genes. In addition, when transplanted into the seminiferous tubule of busulfan-treated mice, SLCs re-located and were maintained in the basal region of the tubule. These results demonstrated that our robust sequential differentiation system produced functional SLCs from mouse ESCs in vitro.

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

“…However, it was hard to isolate adequate eSCs from donor mice for experimental performance. Moreover, the inducing efficiency and purity were not ideal in inducing Sertoli-like cells through RA and Activin A from ESCs ( Bucay et al, 2009 ; Oeda et al, 2013 ; Seol et al, 2018 ). Therefore, we established methods to efficiently deliver non-GM eSLCs via TM4-conditioned medium containing six inducing protein factors, Sry , Sox9 , Sf1 , Wt1 , Gata4 , and Dmrt1 .…”

“…Thus, some research was aimed to provide the solution. Recently, a new approach produced intermediate mesoderm (IM)-derived cells from mouse embryonic stem cells (ESCs) by using some supplements including retinoic acid and Activin A, and then induced the IM-derived cells into embryonic Sertoli-like cells (eSLCs) via some recombinant protein factors including follicle-stimulating hormone (FSH) ( Bucay et al, 2009 ; Oeda et al, 2013 ; Seol et al, 2018 ). However, the differentiation fate and molecular mechanisms in this approach were still unclear.…”

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

“…Then, it is crucial to determine whether the induced eSLCs had similar characteristic and function with eSCs. Some research had isolated mouse Sertoli-like cells by FSHR surface marker [25]. However, no FCM antibody of mouse FSHR on the market or other solid specific surface marker was found for isolating eSCs.…”

“…The main complications of exploring the deriving mechanism of Sertoli cells are as follows: the cells have a great variety in genital ridges and coelomic epithelium, the relevant factors have complicated interaction, the target genes are hard to manipulate in vivo environment, and so on [6,14,18,22,23]. Consequently, recently, some studies provided molecular mechanism through inducing Sertoli-like cells from embryonic stem cells by retiotic acid (RA) treatment, reducing the size of cultured ESC colonies, and some other factors [24,25]. These approaches helped to provide evidences for verifying those established theories.…”

“…In this approach, the cells were manipulated refer to the developmental schedule in vivo. In 0.5-8.5 days, the mESCs were treated by RA and Activin A to induce intermediate mesoderm (IM) [25,32]. In 9.5-10.5 days, Wt1, Gata4, and Sf1 were upregulated through lightswitchable (light-on) transgene system [33][34][35][36].…”