Tailoring Polymer Surface for Long-term Reliable Xenogeneic-free Culture of Intestinal Stem Cells via Regulated Dynamic Migratory Behavior

ABSTRACT: 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.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Stem Cell

DISEASE(S): Disease Free

SUBMITTER: Sun Young Lee  

LAB HEAD: Sung Gap

PROVIDER: PXD064449 | Pride | 2025-07-07

REPOSITORIES: Pride