Recapitulate Organotypic Mesenchyme and Endothelium via Vascularized Lung and Intestine Organoids

Organotypic mesenchyme and endothelium are pivotal during organ development. To generate these context-specific cell types, we established human pluripotent stem cell (hPSC)-derived meso-endoderm spheroids and vascularized primitive gut tube organoids and benchmarked this process via single-cell RNA-seq. Overall design: Day-3 meso-endoderm spheroids, Day-7 vascularized anterior foregut and mid/hindgut, and Day-21 vascularized lung organoid, small intestine, and colon organoids were generated. In Day-3 group, meso-endodermal spheroids receiving four different BMP4 treatment durations were sequenced; in Day-7 group, two vascularized anterior foregut organoids (receive 0 or 1-day BMP4 treatment from day 0-3) and three vascularized mid/hindgut organoids (receive 1, 2, or 3-day BMP4 treatment from day 0-3) were sequenced; in Day 21 group, one vascularized lung organoid, one vascularized small intestine, and one vascularized colon organoid were sequenced.