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Abstract
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Spermatogonial stem cells (SSCs) are a small group of testicular cells located in the basement membrane of
seminiferous tubules and can balance self-renewal and differentiation during spermatogenesis. Our in vitro culture
experiments of mouse SSCs indicated heterogeneity of cultured cells. Highly compact colonies were
observed next to SSC colonies, which we call clump cells. We used immunocytochemical staining to identify
SSCs and somatic cells with VASA and Vimentin antibodies. Subsequently, we compared mRNA expression
levels of VASA, DAZL, PLZF, GFRA1, Lin28, Kit, Myc and Vimentin genes using Fluidigm real-time RTpolymerase
chain reaction in clump cells, SSCs, and testicular stromal cells. To better understand the functions
of selected genes, we created a protein–protein interaction network and performed an enrichment analysis using
different databases. Based on the data collected, we state that clump cells do not express the molecular markers
of SSCs, so we cannot consider them as SSCs; however, we claim that these cells are altered SSCs. The
molecular mechanism of this conversion is still obscure. Therefore, this study can support the analysis of germ
cell development both in vitro and in vivo. In addition, it can be effective in finding new and more efficient
treatments for male infertility.
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