Abstract
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.
Highlights
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
We must recognize that due to the vastness of applications of flow cytometry (FCM) in this area, we have mainly focused on the spermatogenic process itself, leaving aside other applications such as those involving the sorting of spermatozoa
Multiparametric analysis of testicular cell suspensions enables the distinction of various cell types with no need of specific antibodies
Summary
Postnatal testis development has been studied in detail for several species, generating information on the post-partum timing of appearance of specific cell types along the first spermatogenic wave [19,20,21]. Before engaging in spermatogenic cell purification issues, it should be mentioned that many groups have made use of the first spermatogenic wave and employed whole testes from juvenile animals of different ages for downstream molecular studies [22,23,24,25,26,27,28]. These studies have enabled correlations between the observed molecular changes, and the appearance of certain cell types. Single-cell approaches have recently started to be applied to spermatogenesis analysis, as will be elaborated later in this revision
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