Flow cytometry for gene expression studies in Mammalian spermatogenesis

Mammalian spermatogenesis is nowadays still poorly understood at the molecular level, mainly due to the heterogeneous nature of testes, which contain a high number of different cell types, and to the lack of spermatogenic cell culture systems for in vitro studies. As a consequence, the development and/or application of methodological approaches aiming at the enrichment or purification of specific testicular cell types are of great interest and have been addressed by scientists for at least 4 decades. Among the many applications that flow cytometry (FC) has gained since its invention, analysis and sorting of spermatogenic cell populations represent a promising strategy to efficiently overcome testis heterogeneity drawback. Surprisingly, FC has been only rarely used as a preparative method for downstream gene expression studies in specific spermatogenic stages. This work aims to provide an overview of FC for spermatogenic studies including preparation of testicular single cell suspensions, dyes for DNA staining, and our own experience with rodent testis material.