Peptide-based Wnt signal activation enables scalable production of clinical-grade patient-derived intestinal organoids for regenerative cell therapy

Here, we report the successful initiation and expansion of patient-derived intestinal organoids under a GMP-compliant protocol. Culture-, cost-, and time-efficiency improvements were achieved with organoid area-based passaging, one well plate culture, and the incorporation of Wnt activating peptide, PG-008.

The peptide significantly enhanced organoid growth and stabilized patient-patient variability through intestinal stem cell (ISC) enrichment. Single cell RNA sequencing revealed that PG-008 resulted in remarkably pure culture consisting of ISCs and transit amplifying cells, suitable for rapid and consistent expansion. Intriguingly, our GMP-grade intestinal organoids contained LGR5+ ISCs, and injury-induced LGR5- regenerative ISCs, both enriched in peptide culture.