Fate mapping of human P63+ progenitor in vivo by single cell transcriptomic analysis [scRNA_seq]

KRT5+P63+ progenitor cells located in the basal layer of the airway epithelium have been identified as a promising candidate for lung epithelium repair. To gain molecular insights into the fate specification process of human P63+ progenitor cells in the lung, we sorted the transplanted GFP+ cells at different time points using flow cytometry, followed by 10x Genomics single-cell RNA sequencing analysis. Unsupervised clustering of the cells resulted in the identification of 7 distinct populations. We isolated the human P63+ progenitors and transplanted the GFP-labelled P63+ progenitors into the bleomycin-injured mouse lung by intratracheal instillation. Mice were sacrificed at 8 and 35 days post transplantation and their lung tissue were harvest to detect GFP signal by fluorescence stereomicroscope, the region of which was dissect for single cell-RNA-seq (scRNA-seq). We utilized scRNA-seq to trace the fate of transplanted human P63+ progenitors at 8 days and 35 days post transplantation, which revealed cell types differentiated from the transplanted P63+ progenitors.