GEOapplication/xmlftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE301nnn/GSE301802/primaryOK200TranscriptomicsHomo sapiensExpression profiling by high throughput sequencinghttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE301802GEOGSEfalsePeptide-based Wnt signal activation enables scalable production of clinical-grade patient-derived intestinal organoids for regenerative cell therapyDespite the clinical potential of intestinal organoids as a resource for regenerative cell therapy and bioengineering, the lack of reliable clinical-grade cultures has hampered further development. 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. Our study establishes clinical-grade intestinal organoids for further application, and a model facilitating in vitro investigations into regenerative stem cell induction and intestinal regeneration.2026/04/01GSE301802GSM9090003GSM909000424676301802Homo sapiens[41906165]