Analysis of gene expression heteroegeneity in adult undifferentiated spermatogonia [single-cell RNA-Seq]

The undifferentiated spermatogonial population of mouse testis is functionally heterogeneous and contains stem cells and committed progenitor cells. However, gene expression patterns marking these distinct cell fractions are poorly defined. Therefore, our aim was to profile gene expression of undifferentiated cells by RNA-Seq at the single cell level. Undifferentiated cells were therefore isolated from adult mouse testes based on expression of a Plzf/Zbtb16 gene reporter, which marks both stem and progenitor spermatogonia. Isolated cells were then processed using the 10X Chromium system and analysed by RNA-Seq. Our goal was to characterize distinct subsets of undifferentiated spermatogonia based on gene expression patterns. Undifferentiated spermatogonia were independently isolated from 2 adult Plzf-mCherry/CreER reporter adult mice and samples processed independently.小鼠睾丸未分化精原细胞群体在功能上具有异质性，包含干细胞与定向祖细胞。然而，用以区分这些不同细胞亚群的基因表达特征仍未得到充分阐明。因此，本研究旨在通过单细胞RNA测序（RNA-Seq）技术分析未分化细胞的基因表达谱。研究人员基于可标记干细胞和祖代精原细胞的Plzf/Zbtb16基因报告基因的表达，从成年小鼠睾丸中分离未分化细胞。随后利用10X Chromium系统对分离得到的细胞进行处理，并通过RNA测序进行分析。本研究的目标是基于基因表达特征对未分化精原细胞的不同亚群进行鉴定与表征。研究人员分别从2只成年Plzf-mCherry/CreER报告基因小鼠中独立分离未分化精原细胞，并对样本进行独立处理。