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

• A GMP-compliant protocol using Type-I collagen achieves an 82% establishment rate for patient-derived colonic organoids from 60 patients. • Incorporation of the Wnt-activating peptide PG-008 enhances organoid growth and reduces patient-to-patient variability by enriching intestinal stem cells. • Single-cell RNA sequencing reveals that PG-008 yields a remarkably pure culture of LGR5+ and LGR5− regenerative intestinal stem cells. • This scalable, clinical-grade organoid platform supports applications in autologous transplantation and bioengineering for gastrointestinal regenerative therapy.