Generation of human iPS cell-derived mesenchymal cells capable of forming alveolar organoids [bulkRNA-Seq]

We developed a method of generating human induced pluripotent stem cell (iPSC)-derived mesenchymal cells (iMES) that were able to induce AT1 and AT2 epithelial lineage cells in organoids. Transcriptomic analysis of human fetal lung fibroblasts (HFLF), human dermal fibroblasts (HDF), and their corresponding iMES revealed that iMES harbor characteristics of lung distal tip mesenchyme. Overall design: We stepwise differentiated SFTPC-GFP reporter iPSCs into lung progenitor cells and they were co-cultured with HFLF, HDF or their corresponding iMES in the organoid differentiation system. Each mesenchyme in pre-3D culture condition was collected and total RNA was extracted, respectively. In addition, each mesenchyme in post-3D culture condition was isolated by fluorescence-activated cell sorting (FACS) after gating for EPCAM negative cells derived from the alveolar organoids at P0 and total RNA was extracted as well. The transcriptomes of these cells were sequenced with paired-end reads (100-bp) on an Illumina Novaseq 6000 instrument.