Department of Molecular Biology, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), 11600 Montevideo, Uruguay.
Biochemistry-Molecular Biology, Facultad de Ciencias, Universidad de la República (UdelaR), 11400 Montevideo, Uruguay.
Int J Mol Sci. 2021 Jan 25;22(3):1151. doi: 10.3390/ijms22031151.
Mammalian testes are very heterogeneous organs, with a high number of different cell types. Testicular heterogeneity, together with the lack of reliable in vitro culture systems of spermatogenic cells, have been an obstacle for the characterization of the molecular bases of the unique events that take place along the different spermatogenic stages. In this context, flow cytometry has become an invaluable tool for the analysis of testicular heterogeneity, and for the purification of stage-specific spermatogenic cell populations, both for basic research and for clinical applications. In this review, we highlight the importance of flow cytometry for the advances on the knowledge of the molecular groundwork of spermatogenesis in mammals. Moreover, we provide examples of different approaches to the study of spermatogenesis that have benefited from flow cytometry, including the characterization of mutant phenotypes, transcriptomics, epigenetic and genome-wide chromatin studies, and the attempts to establish cell culture systems for research and/or clinical aims such as infertility treatment.
哺乳动物的睾丸是非常异质的器官,具有大量不同的细胞类型。睾丸的异质性,加上缺乏可靠的生精细胞体外培养系统,一直是描述不同生精阶段独特事件的分子基础的障碍。在这种情况下,流式细胞术已成为分析睾丸异质性和纯化阶段特异性生精细胞群体的宝贵工具,无论是用于基础研究还是临床应用。在这篇综述中,我们强调了流式细胞术对于哺乳动物精子发生分子基础研究进展的重要性。此外,我们还提供了一些受益于流式细胞术的精子发生研究的不同方法的例子,包括突变表型的特征描述、转录组学、表观遗传学和全基因组染色质研究,以及建立细胞培养系统以用于研究和/或临床目的(如治疗不育症)的尝试。