Department of Comparative Biosciences, College of Veterinary Medicine, University of Illinois Urbana Champaign, Urbana, Illinois.
Mol Reprod Dev. 2020 Aug;87(8):857-870. doi: 10.1002/mrd.23402. Epub 2020 Aug 2.
Sertoli cells play a vital role in spermatogenesis by offering physical and nutritional support to the differentiating male germ cells. They form the blood-testis barrier and secrete growth factors essential for germ cell differentiation. Sertoli cell primary cultures are critical for understanding the regulation of spermatogenesis; however, obtaining pure cultures has been a challenge. Rodent Sertoli cell isolation protocols do not rule out contamination by the interstitial or connective tissue cells. Sertoli cell-specific markers could be helpful, but there is no consensus. Vimentin, the most commonly used marker, is not specific for Sertoli cells since its expression has been reported in peritubular myoid cells, mesenchymal stem cells, fibroblasts, macrophages, and endothelial cells, which contaminate Sertoli cell preparations. Markers based on transcription and growth factors also have limitations. Thus, the impediment to obtaining pure Sertoli cell cultures pertains to both the method of isolation and marker usage. The aim of this review is to discuss improvements to current methods of rodent Sertoli cell primary cultures, assess the properties of prepubertal versus mature Sertoli cell cultures, and propose steps to improve cellular characterization. Potential benefits of using contemporary approaches, including lineage tracing, specific cell ablation, and RNA-seq for obtaining Sertoli-specific transcript markers are discussed. Evaluating the specificity and applicability of these markers at the protein level to characterize Sertoli cells in culture would be critical. This review is expected to positively impact future work using primary cultures of rodent Sertoli cells.
支持细胞在精子发生中起着至关重要的作用，为分化中的雄性生殖细胞提供物理和营养支持。它们形成血睾屏障并分泌对生殖细胞分化至关重要的生长因子。支持细胞原代培养对于理解精子发生的调节至关重要；然而，获得纯培养一直是一个挑战。啮齿动物支持细胞分离方案不能排除间质或结缔组织细胞的污染。支持细胞特异性标志物可能会有所帮助，但目前尚无共识。波形蛋白是最常用的标志物，但它并不特异，因为它的表达已在小管周肌样细胞、间充质干细胞、成纤维细胞、巨噬细胞和内皮细胞中报道过，这些细胞会污染支持细胞的制备物。基于转录和生长因子的标志物也存在局限性。因此，获得纯支持细胞培养物的障碍既涉及分离方法，也涉及标志物的使用。本综述的目的是讨论改进目前的啮齿动物支持细胞原代培养方法，评估青春期前和成熟支持细胞培养物的特性，并提出改善细胞特征的步骤。讨论了使用当代方法的潜在益处，包括谱系追踪、特定细胞消融和 RNA-seq 以获得支持细胞特异性转录标志物。评估这些标志物在蛋白质水平上对培养物中支持细胞的特异性和适用性至关重要。本综述有望对未来使用啮齿动物支持细胞原代培养的工作产生积极影响。
