10x Genomics Drop-seq single-cell RNA-seq of isolated Adult ID4-EGFP mouse spermatogonia, spermatocytes, spermatids & steady-state spermatogenic cells

To reveal distinct transcriptomes associated with various spermatogenic cells, including spermatogonial stem cells and all of their subsequent progeny, single-cell transcriptomes from Adult ID4-EGFP+ spermatogonia (sorted for brightest or dimmest), StaPut-enriched spermatocytes and spermatids, or unselected steady-state spermatogenic cells were used for 10x Genomics analysis. The GFP-bright and dim phenotypes exhibit distinct fates when assayed by transplantation, with ID4-EGFPbright cells highly enriched for SSCs, and ID4-EGFPdim cells enriched for progenitors. We used the 10x Genomics Chromium (Drop-Seq) to perform single-cell RNA-seq Unselected or sorted Adult ID4-EGFP+ spermatogonia (sorted separately as EGFP-bright or EGFP-dim), three replicate preparations of steady-state unselected spermatogenic cells, and isolated adult spermatocytes and spermatids were used for this study. Data are from a total of 17,491 cells.为揭示与各类生精细胞（包括精原干细胞(Spermatogonial Stem Cells, SSCs)及其所有后续子代细胞）相关的独特转录组特征，本研究采用成年ID4-EGFP阳性精原细胞（按荧光亮度最高或最低分选）、StaPut富集的精母细胞与精子细胞，以及未分选的稳态生精细胞的单细胞转录组进行10x Genomics分析。GFP高亮度与低亮度表型的细胞在移植实验中展现出截然不同的命运：ID4-EGFP高亮度细胞高度富集精原干细胞，而ID4-EGFP低亮度细胞则富集于祖细胞群体。本研究使用10x Genomics Chromium（Drop-Seq技术平台）开展单细胞RNA测序，所用样本包括未分选或分别按EGFP亮度分选的成年ID4-EGFP阳性精原细胞、三份生物学重复的稳态未分选生精细胞，以及分离得到的成年精母细胞与精子细胞。本研究数据集共包含17491个细胞。