Single-cell RNA-seq of MEFs transduced with 4 factors (Nkx2-1, Foxa1, Foxa2, Gata6), 14 days after the transduction

Single cell RNA-seq analyses of MEFs obtained from GFP/SP-C transgenic mice on CBA/Ca x C57BL/6 mixed background were transduced sets of 4 combination factors (Nkx2-1/Foxa1/Foxa2/Gata6) into induced pulmonary epithelial cell-like cells 14 days after the transduction. Abstract is as follows: pneumocyte differentiation has been achieved previously using embryonic stem cells or induced pluripotent stem cells. However, direct reprograming of somatic cells into pulmonary epithelial cells has not been achieved. Here, we report that a combination of three or four transcription factors (i.e., Nkx2-1, Gata6, and [Foxa1 and/or Foxa2]) directly reprogrammed mouse tail-tip fibroblast or embryonic fibroblast into differentiated induced pulmonary epithelial cell-like cells (iPUL cells). iPUL cells had a global gene expression profile similar to various kinds of pulmonary epithelial cells. Some iPUL cells showed lamellar body-like structures and expressed SP-C, consistent with the features of alveolar epithelial type II cells. Interestingly, intratracheal administration of iPUL cells rescued influenza virus-induced acute lung injury in mice. These findings demonstrate that functional pulmonary epithelial cell-like cells can be directly reprogrammed from differentiated somatic cells by defined factors. Reprograming of fibroblasts might provide a source of pulmonary epithelial cells for regenerative medicine. We generated pulmonary epithelial-like cells transduced 4 factors (Nkx2-1, Foxa1, Foxa2, Gata6) from mouse embryonic fibroblast and the cells were subjected to scRNA-Seq on the 10X Chromium System (10X Genomics, Pleasanton, CA) with the v3 chemistry. We sequenced (Read1=28 bp, Read2=98 bp and Readi7=8 bp) on a Illumina NovaSeq 6000 (Illumina, San Diego, CA).