Single cell transcriptome analysis of distal adult murine lung airway epithelium

To comprehensively study the heterogeneity within distal airway epithelium, we performed single cell transcriptomic analysis of the naive lung samples. Our analysis reveals that distal lung airway epithelial cells are highly heterogenous. We employed supervised clustering to identify a rare group of cells with transcriptional similiarities to embryonic distal lung bud tip progenitors. This approach identified a quiescent progenitor population of airway cells that accounted for all in vitro regenerative activity. Furthermore, these cells were also required for in vivo transplantation, reconstitution of alveoli, and functional recovery of injured mice. These findings indicate that small subpopulations of specialized stem/progenitors are required for effective lung regeneration and could potentially be used as therapeutic adjuncts after major lung injury. Overall design: We performed single cell transcriptomic sequencing of flow purified Epcam+/Itgb4+/CD200+ cells via 10X Genomics platform为全面探究远端气道上皮的细胞异质性，我们对未处理肺组织样本开展了单细胞转录组分析。本分析结果显示，远端肺气道上皮细胞具有高度异质性。我们采用有监督聚类（supervised clustering）技术，鉴定出一群转录特征与胚胎远端肺芽尖祖细胞高度相似的稀有细胞亚群。该方法筛选得到一类气道静息祖细胞群，此类细胞具备全部体外再生活性。此外，这群细胞亦是体内移植、肺泡重建以及损伤小鼠肺功能恢复所必需的。本研究表明，少量特化干/祖细胞亚群是高效肺再生过程的必需组分，有望成为重度肺损伤后的治疗辅助手段。实验设计概述：我们通过10X基因组学（10X Genomics）平台，对经流式分选纯化的Epcam+/Itgb4+/CD200+细胞开展了单细胞转录组测序。