Reproductive Biology Laboratory, Center for Reproductive Medicine, Amsterdam UMC, Amsterdam Reproduction & Development Research Institute, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands.
Department of Epidemiology & Data Science, Amsterdam UMC, Amsterdam Public Health Research Institute, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands.
Int J Mol Sci. 2020 Nov 4;21(21):8269. doi: 10.3390/ijms21218269.
Autologous spermatogonial stem cell transplantation is an experimental technique aimed at restoring fertility in infertile men. Although effective in animal models, in vitro propagation of human spermatogonia prior to transplantation has proven to be difficult. A major limiting factor is endogenous somatic testicular cell overgrowth during long-term culture. This makes the culture both inefficient and necessitates highly specific cell sorting strategies in order to enrich cultured germ cell fractions prior to transplantation. Here, we employed RNA-Seq to determine cell type composition in sorted integrin alpha-6 (ITGA6+) primary human testicular cells ( = 4 donors) cultured for up to two months, using differential gene expression and cell deconvolution analyses. Our data and analyses reveal that long-term cultured ITGA6+ testicular cells are composed mainly of cells expressing markers of peritubular myoid cells, (progenitor) Leydig cells, fibroblasts and mesenchymal stromal cells and only a limited percentage of spermatogonial cells as compared to their uncultured counterparts. These findings provide valuable insights into the cell type composition of cultured human ITGA6+ testicular cells during in vitro propagation and may serve as a basis for optimizing future cell sorting strategies as well as optimizing the current human testicular cell culture system for clinical use.
自体精原干细胞移植是一种旨在恢复不育男性生育能力的实验技术。尽管在动物模型中有效，但在移植前体外培养人类精原细胞已被证明具有挑战性。一个主要的限制因素是长期培养过程中内源性体睾丸细胞的过度生长。这使得培养效率低下，并且需要高度特异性的细胞分选策略，以便在移植前富集培养的生殖细胞分数。在这里，我们使用 RNA-Seq 来确定长达两个月培养的分选整合素 alpha-6 (ITGA6+) 原代人睾丸细胞（= 4 个供体）中的细胞类型组成，使用差异基因表达和细胞去卷积分析。我们的数据和分析表明，与未培养的细胞相比，长期培养的 ITGA6+睾丸细胞主要由表达小管周肌样细胞、（祖）莱迪希细胞、成纤维细胞和间充质基质细胞标志物的细胞组成，而精原细胞的比例有限。这些发现为体外培养过程中培养的人 ITGA6+睾丸细胞的细胞类型组成提供了有价值的见解，并可能为优化未来的细胞分选策略以及优化当前的人睾丸细胞培养系统用于临床用途提供基础。
