Single cell RNA sequencing of distal lund epithelial cells generated from human induced pluripotent stem cells

We perform transcriptomic profiling of a directed differentiation protocol for generating distal lung epithelial cells and alveolar type II epithelial cells (AEC2s) from pluripotent stem cells (PSCs) using bi-fluorescently targeted iPSC cell line BU3-NGST. On day 33 of distal lung differentiation, we sorted SFTPCtdTomato/NKX2-1GFP double positive cells and NKX2-1GFP single positive cells for Fluidigm single cell RNA sequencing analysis. We find canonical AEC2s genes expressed in both sorted populations but at a higher level in the SFTPCtdTomato/NKX2-1GFP double positive population. We also find genes associated with cellular proliferation to be enriched in the NKX2-1GFP single positive population, suggestive of a less mature but more proliferative subset of PSC-derived AEC2s. Overall design: Global single cell transcriptomic analysis of day 33 PSC-derived distal lung epithelial cell populations identified by NKX2-1GFP and SFTPCtdTomato reporters.

本研究针对利用双荧光靶向标记诱导多能干细胞（induced pluripotent stem cell, iPSC）系BU3-NGST，从多能干细胞（pluripotent stem cells, PSCs）定向分化获取远端肺上皮细胞与II型肺泡上皮细胞（alveolar type II epithelial cells, AEC2s）的实验方案，开展转录组谱分析。在远端肺定向分化第33天时，我们分别分选了SFTPCtdTomato/NKX2-1GFP双阳性细胞与NKX2-1GFP单阳性细胞，用于Fluidigm单细胞RNA测序分析。研究发现，两类分选细胞群中均有经典AEC2s特征基因表达，但SFTPCtdTomato/NKX2-1GFP双阳性细胞群中的表达水平更高。同时，我们观察到与细胞增殖相关的基因在NKX2-1GFP单阳性细胞群中显著富集，提示该细胞群为一类成熟度较低但增殖活性更强的多能干细胞来源AEC2s亚群。本研究的总体设计为：基于NKX2-1GFP与SFTPCtdTomato报告基因标记的、分化第33天的多能干细胞来源远端肺上皮细胞群的全局单细胞转录组分析。