The Human Testis Cell Atlas via Single-cell RNA-seq (KIT+ spermatogonia ATAC-seq data set)

Human adult spermatogenesis involves a balance of spermatogonial stem cell self renewal and differentiation, alongside complex germline-niche interactions. To better understand, we performed single cell RNA sequencing of ~7000 testis cells from three healthy men of peak reproductive age. Our analyses revealed multiple distinctive transcriptional ‘states’ of self-renewing and differentiating spermatogonia, the cellular stages of gametogenesis, five niche cells (Leydig, Myoid, Sertoli, Endothelial, macrophage) and insights into germline-niche communication. Spermatogenesis was reconstructed computationally, which identified sequential coding, noncoding, and repeat-element transcriptional signatures. A new, developmentally early and likely quiescent spermatogonial state is identified (GFRA1-/ETV5-/ID4+/UTF1+/FGFR3+). Notably, certain epigenetic features combined with nascent transcription analyses suggest considerable plasticity within certain spermatogonial populations/states. Key findings were validated via RNA and protein staining. Taken together, we provided the first “Cell Atlas” of the adult human testis, and provide multiple new insights into germ cell development and germ cell – niche interaction.

人类成年精子发生过程需要维持精原干细胞（spermatogonial stem cell）自我更新与分化的动态平衡，同时伴随复杂的生殖细胞微环境（germline-niche）相互作用。为深入解析这一生物学过程，我们对3名生育旺盛期健康男性的约7000个睾丸细胞开展了单细胞RNA测序（single cell RNA sequencing）。分析结果揭示了自我更新型与分化型精原细胞（属于配子发生（gametogenesis）的细胞阶段）的多种独特转录状态，同时鉴定出5类微环境细胞：莱迪希细胞（Leydig）、肌样细胞（Myoid）、支持细胞（Sertoli）、内皮细胞（Endothelial）与巨噬细胞（macrophage），并为生殖细胞-微环境通信机制提供了新见解。我们通过计算手段重构了精子发生过程，鉴定出一系列连续的编码基因、非编码RNA及重复元件的转录特征。本研究还发现了一种新的发育早期且大概率处于静息状态的精原细胞状态（GFRA1-/ETV5-/ID4+/UTF1+/FGFR3+）。值得注意的是，结合表观遗传（epigenetic）特征与新生转录（nascent transcription）分析结果，提示部分精原细胞群体/状态具有显著的可塑性。核心研究发现均通过RNA与蛋白染色实验得到了验证。综上，本研究构建了首个成人睾丸细胞图谱（Cell Atlas），并为生殖细胞发育及生殖细胞-微环境相互作用领域提供了多项全新认知。

搜集汇总

背景与挑战

背景概述

该数据集基于三名健康成年男性约7000个睾丸细胞的单细胞RNA测序，构建了人类睾丸细胞图谱，揭示了精子发生过程中的多个转录状态和细胞类型，包括新的精原细胞状态和生态位细胞相互作用。它提供了对生殖细胞发育和表观遗传可塑性的新见解，并通过实验验证了关键发现，是首个成人睾丸单细胞水平的全面图谱。