Single cell sequencing facilitates quantitative analysis of transcriptomes of proliferative alveolar epithelial type 2 (AT2) cells in adult mouse lungs

Purpose: The goals of this study are to generate transcriptome profiling at single cell level (scRNA-seq) of proliferative AT2 cells Methods: Lungs of 6- to 10-week-old SPC-EGFP, Ki67-creRT2, Rosa26-tdTomato mice before and 4 days after Streptococcus pneumoniae infection-induced lung injury were dissociated using dispase digesting method. Proliferative AT2 cells (GFP+/tdTomato+) and non-proliferative AT2 cells (GFP+/tdTomato-) were sorted using a BD influx LSRII. AT2 cell mRNA profiles were generated by scRNA-seq. Sequencing was performed on an Illumina Nextseq 500 system operated by the Next Generation Sequencing at GENEWIZ using the HiSeq 2x150 bp high output sequencing kit.  The scRNAseq data was processed by Cell Ranger (v.3.0.2, 10X Genomics) followed by Seurat suite (version 4.0.1). Results: Our results show a distinctive transcriptome profile of proliferative AT2 cells in adult mouse lungs Conclusions: Our study represents the first detailed analysis of transcriptomes of proliferative and non-proliferative AT2 cells in adult mouse lungs generated by scRNA-seq technology. mRNA profiles of proliferative AT2 cells (GFP+/tdTomato+) and non-proliferative AT2 cells (GFP+/tdTomato-) in adult mouse lungs at homeostasis and after injury