Abstract
Sertoli cells are main players in the male gonads development and their study may shed light on 46,XY disorders of sex development (DSD). Mature primary Sertoli cells are incapable of proliferating in prolonged in vitro cultures and the available Sertoli cell models have several limitations since they derive from mouse or human cancer tissues. We differentiated human fibroblasts (HFs)-derived induced pluripotent stem cells into Sertoli-like cells (SLC) and, in order to characterize this new Sertoli cell model, we performed gene expression analyses by NextGeneration Sequencing techniques. This approach revealed that our putative SLC have reduced expression of pluripotency markers and expressed Sertoli cell markers such as SRY-Related HMG-Box 9 (SOX9), vimentin (VIM), and claudin-11 (CLDN-11). More in detail, the transcriptional profile analysis suggested that these cells are in an early stage of Sertoli cells maturation. Harnessing the power of induced pluripotent stem cells, we were able to generate SLC that show genetic and functional similarities to human Sertoli cells (HSerCs). SLC could become an excellent source of patient-specific Sertoli cells that could be of paramount benefit for both basic research and personalized medicine in sex development and reproductive medicine.
Highlights
The normal development of a male individual depends on the development of a functional testis where Sertoli cells (SCs) play a pivotal role
SNL Feeder cells (CBA-136, Cell Biolab, San Diego, CA, United States) were cultured in DMEM supplemented with 10% FBS, 1% Pen/Strep and 0.1 mM MEM Non-essential Amino Acids (NEAA), when a confluence of about 96% was reached, SNL feeder cells were treated with Mitomycin C (MMC) and seeded in Matrigel coated plates as a feeder layer for human fibroblasts (HFs) de-differentiation
In vivo staining with alkaline phosphatase (AP) live stain identified the colonies as potential induced-pluripotent stem cells (iPSCs)
Summary
The normal development of a male individual depends on the development of a functional testis where Sertoli cells (SCs) play a pivotal role. SCs cells are the first cell type to differentiate in the bipotential fetal male gonad, in turn enabling testicular cord formation and controlling the differentiation and function of Leydig cells (Racine et al, 1998). Leydig cells, for their part, secrete testosterone and play important roles in downstream masculinization events (Barsoum and Yao, 2010)
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