Dataset Information

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

ABSTRACT: Despite 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.

ORGANISM(S): Homo sapiens

PROVIDER: GSE301802 | GEO | 2026/04/01

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Regenerative therapy employing intestinal stem cells (ISCs) holds a great promise for reliable epithelial restoration. However, achieving this goal requires scalable and robust, but fully defined culture platforms that eliminate the use of xenogeneic components while preserving stem cell function. Most existing systems usually rely on the use of Matrigel, complicating clinical translation and limiting mechanistic understanding of stem cell-material interactions. In this study, a poly(ethyleneglycoldimethacrylate) (pEGDMA)-based synthetic culture surface is precisely refined through N2 plasma treatment thereon, enabling tailored modulation of surface properties. This modification enhances wettability of the polymer surface and introduces N-containing functional groups, resulting in the PoLymer-coated Ultra-stable Surface (PLUS) that significantly improves ISC attachment under xenogeneic-free conditions. Remarkably, PLUS maintains its ISC-supportive function even after three years of ambient storage. Gene expression analysis and comparative proteomic profiling clearly reveal the upregulation of factors involved in actin dynamics and cytoskeletal reorganization, indicating a structural basis for enhanced colony expansion. Consistently, colonies on PLUS exhibit a broader migratory range and migrate 1.8-fold faster during random movement than those on pristine pEGDMA. Functional validation using small-molecule perturbation assays confirms the involvement of cytoskeletal remodeling by exquisitely mediating ISC-substrate interaction. Furthermore, PLUS enables progressive wound closure via dynamic migration by up to 46.7% within 144 h through actin-dependent mechanisms, supporting the facilitated epithelium regeneration. By providing a long-term reliable bioactive surface that sustains stemness and regenerative capacity of ISCs, PLUS engages cytoskeletal machinery as a central mediator of ISC-substrate interaction, in part by promoting actin-dependent cytoskeletal reorganization, positioning it as a scalable, translational platform for intestinal stem cell-based regenerative medicine.

Dataset Information

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

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets