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 enables clinical-grade colonic organoid generation from endoscopic biopsies with an 82% establishment rate across 60 patients. • The Wnt-activating peptide PG-008 significantly enhances organoid growth and stabilizes patient-to-patient variability by enriching intestinal stem cells (ISCs). • Single-cell RNA sequencing demonstrates that PG-008 yields a remarkably pure ISC population, outperforming conventional recombinant WNT3A culture. • The scalable, cost-effective culture strategies (area-based passaging, one-well plate) facilitate mass production of clinical-grade organoids for regenerative cell therapy.