Differentiation of Peritubular Myoid‐Like Cells from Human Induced Pluripotent Stem Cells

• Published in: Advanced biology 2023-07, Vol.7 (7), p.e2200322-n/a
• Main Authors: Robinson, Meghan, Haegert, Anne, Li, Yen‐Yi, Morova, Tunc, Zhang, Angelina Yuan Yuan, Witherspoon, Luke, Hach, Faraz, Willerth, Stephanie M., Flannigan, Ryan
• Format: Article
• Language: English
• Subjects: cell culture; cell differentiation; Cell Differentiation - genetics; human induced pluripotent stem cells; Humans; Induced Pluripotent Stem Cells; infertility; Infertility, Male - metabolism; Leydig cells; Male; peritubular myoid cells; platelet‐derived growth factor; Spermatogenesis - genetics; Testis - metabolism
• ISSN: 2701-0198; 2701-0198
• DOI: 10.1002/adbi.202200322

Infertility affects 10–15% of couples, with half attributed to male factors. An improved understanding of the cell‐type‐specific dysfunction contributing to male infertility is needed to improve available therapies; however, human testicular tissues are difficult to obtain for research purposes. To overcome this, researchers have begun to use human induced pluripotent stem cells (hiPSCs) to generate various testis‐specific cell types in vitro. Peritubular myoid cells (PTMs) are one such testicular cell type that serves a critical role in the human testis niche but, to date, have not been derived from hiPSCs. This study set forth to generate a molecular‐based differentiation method for deriving PTMs from hiPSCs, mirroring in vivo patterning factors. Whole transcriptome profiling and quantitative polymerase chain reaction (qPCR) show that this differentiation method is sufficient to derive cells with PTM‐like transcriptomes, including upregulation of hallmark PTM functional genes, secreted growth and matrix factors, smooth muscle, integrins, receptors, and antioxidants. Hierarchical clustering shows that they acquire transcriptomes similar to primary isolated PTMs, and immunostaining shows the acquisition of a smooth muscle phenotype. Overall, these hiPSC‐PTMs will allow in vitro study of patient‐specific PTM development and function in spermatogenesis and infertility. Studying human testis development and spermatogenesis has been limited to the ability to enrich pure populations of testicular cell types. Here, a novel protocol is described using human‐induced pluripotent stem cells to engineer testicular myoid‐like cells. These cells play a critical role in supporting spermatogenesis and may serve as a tool to model normal and diseased human in vitro spermatogenesis.