Comparison of iPSC-derived alveolar epithelial cell culture methods

We established a novel alveolar epithelial culture method, called "On-Gel" culture. To characterize the "On-Gel" culture, we compared each transcriptome of the cultured cells in "On-Gel", fibroblast dependent-alveolar organoids (FD-AO) and fibroblast-free alveolar organoids (FF-AO) and their progenitor cells (CPMhigh Lung Progenitors). Overall design: Human iPSCs were differentiated into lung progenitor cells. After isolated with anti-Carboxypepdidase M (CPM) antibody, CPMhigh Lung Progenitors were co-cultured with human fetal lung fibroblasts (HFLFs) to generate FD-AOs in which surfactant protein C (SPC)-positive cells appeared (Passage 0: P0). SPC-positive cells were isolated by FACS and cultured in each condition for generating FD-AOs, FF-AOs, or "On-Gel" spheroids (P1). EpCAM-positive cells of FD-AOs derived from B2-3 SFTPC-GFP reporter iPSCs were isolated by MACS and the transcriptomes of P1 epithelial cells in each culture method were analyzed.