College of Life Sciences, Key Laboratory of Animal Reproduction and Biotechnology in Universities of Shandong, Qingdao Agricultural University, Qingdao, 266109, China.
Department of Animal Sciences, Auburn University, Auburn, AL, 36849, USA.
Stem Cell Res Ther. 2023 Feb 3;14(1):17. doi: 10.1186/s13287-023-03243-5.
Many laboratories have described the in vitro isolation of multipotent cells with stem cell properties from the skin of various species termed skin-derived stem cells (SDSCs). However, the cellular origin of these cells and their capability to give rise, among various cell types, to male germ cells, remain largely unexplored.
SDSCs were isolated from newborn mice skin, and then differentiated into primordial germ cell-like cells (PGCLCs) in vitro. Single-cell RNA sequencing (scRNA-seq) was then applied to dissect the cellular origin of SDSCs using cells isolated from newborn mouse skin and SDSC colonies. Based on an optimized culture strategy, we successfully generated spermatogonial stem cell-like cells (SSCLCs) in vitro.
Here, using scRNA-seq and analyzing the profile of 7543 single-cell transcriptomes from newborn mouse skin and SDSCs, we discovered that they mainly consist of multipotent papillary dermal fibroblast progenitors (pDFPs) residing in the dermal layer. Moreover, we found that epidermal growth factor (EGF) signaling is pivotal for the capability of these progenitors to proliferate and form large colonies in vitro. Finally, we optimized the protocol to efficiently generate PGCLCs from SDSCs. Furthermore, PGCLCs were induced into SSCLCs and these SSCLCs showed meiotic potential when cultured with testicular organoids.
Our findings here identify pDFPs as SDSCs derived from newborn skin and show for the first time that such precursors can be induced to generate cells of the male germline.
许多实验室已经描述了从各种物种的皮肤中分离具有干细胞特性的多能细胞，这些细胞被称为皮肤来源的干细胞（SDSCs）。然而，这些细胞的细胞起源及其在各种细胞类型中产生雄性生殖细胞的能力在很大程度上仍未得到探索。
从新生小鼠的皮肤中分离 SDSCs，然后在体外将其分化为原始生殖细胞样细胞（PGCLCs）。然后应用单细胞 RNA 测序（scRNA-seq）来剖析 SDSCs 的细胞起源，使用从新生小鼠皮肤和 SDSC 集落中分离的细胞。基于优化的培养策略，我们成功地在体外产生了精原干细胞样细胞（SSCLCs）。
在这里，我们使用 scRNA-seq 分析了来自新生小鼠皮肤和 SDSCs 的 7543 个单细胞转录组的图谱，发现它们主要由多能性乳突真皮成纤维细胞祖细胞（pDFPs）组成，这些祖细胞位于真皮层。此外，我们发现表皮生长因子（EGF）信号对于这些祖细胞在体外增殖和形成大集落的能力至关重要。最后，我们优化了从 SDSCs 中高效产生 PGCLCs 的方案。此外，PGCLCs 被诱导为 SSCLCs，当与睾丸类器官共培养时，这些 SSCLCs 显示出减数分裂潜能。
我们的研究结果确定了 pDFPs 为来自新生皮肤的 SDSCs，并首次表明这些前体能被诱导产生雄性生殖细胞。
