RNAseq of iPSC derived pulmonary organoids by co-culturing endodermal and mesodermal progenitors
RNAseq of iPSC derived pulmonary organoids by co-culturing endodermal and mesodermal progenitors
4 iPSC lines used are: SOJD3, SEUR7, WT02C3 and WT02C9. nto anterior foregut endoderm and mesoderm progenitors. The two population of cells were imbedded in Matrigel and co-cultured in an insert of a 12 mm Transwell that was emerged in DMEM base medium supplemented with human serum replacement 3, BMP4, FGF2, FGF7, and FGF10. The medium was changes every 2-3 days and the pulmonary organoids spontaneously formed within 14 days of the co-culture. The pulmonary organoids were first treated with Corning cell recovery solution (Corning, Deeside, UK) to recover cells before nucleic RNA extraction. The induced pluripotent stem cells were cultured in Essential 8 Medium (Thermo Fisher Scientific) The pulmonary organoids were first treated with Corning cell recovery solution (Corning, Deeside, UK) to recover cells, and then the total RNAs were extracted using TRIZOL and phenol-chloroform. Total RNA from iPSCs was extracted using the RNeasy Mini Kit (Qiagen). Illumina TruSeq Stranded mRNA kit, according to the manufacturerâ€™s protocol.