In vitro maturation of human pluripotent stem cell-derived intestinal organoids

Human intestinal organoids (hIOs) derived from human pluripotent stem cells (hPSCs) have immense potential as a source of intestines. However, due to problems obtaining mature adult-like cells from hPSCs, effective strategies for the maturation of hIOs with improved functionalities must be developed. We established conditions that enable the in vitro maturation of hIOs derived from hPSCs, recapitulating the essential features of intestinal tissue. The maturity of the hPSC-derived hIOs was enhanced by the in vitro maturation in our co-culture system, evidenced by transcriptional changes observed in our microarray data. Global transcription of in vitro-matured hIOs is highly similar to that of adult human small intestine (hSI). Compared to control hIOs, in vitro-matured hIOs exhibited significant increases in the expression of intestinal maturation markers, major transporters, and key enzymes for drug absorption and first-pass metabolism. Changes in the gene expression profile of undifferentiated human pluripotent stem cells (hPSC), definitive endoderm cells (DE), hindgut cells (HG), hIOs (p0), hIOs (p2), in vitro matured hIOs and adult human small intestine (hSI). At least three independent experiments were performed for each condition.