Mechanically Stable and Reactive Oxygen Species-Responsive Hydrogel-Coated Cell Culture Plate for Enhanced Stem Cell Activation

Stem cells have a distinctive capacity for self-renewal and the potential to develop into multiple specialized cell types, while their paracrine signaling mechanisms play a pivotal role in modulating the microenvironment to support regeneration. However, conventional cell culture systems often fail to sufficiently stimulate these stem cell functions, presenting a significant barrier to their therapeutic application. Here, we developed a novel hydrogel-coated cell culture platform using gelatin reinforced with tannic acid (GT) and functionalized with a Pluronic F68 (PF68)-Chlorin e6 (Ce6) conjugate (GTPXC). Upon controlled laser irradiation, the GTPXC-coated cell culture platform effectively generates reactive oxygen species (ROS), which acts as a biochemical stimulus to significantly enhance stem cell proliferation, viability, and paracrine signaling activity. To validate the platform, we conducted comprehensive structural and rheological analyses, which confirmed the successful integration of tannic acid and PF68-Ce6 conjugates into the gelatin matrix. The GTPXC exhibited enhanced mechanical stability, essential for maintaining a conducive environment for prolonged stem cell culture. Biological evaluations, including cellular proliferation test, exosome quantification, cell cycle assay, and characterization of differentiation potential, demonstrated the ability of the GTPXC-coated platform to activate stem cells and improve their functional efficacy compared to conventional culture systems. This ROS-responsive GTPXC-coated cell culture platform presents a promising strategy for improving stem cell functionality in regenerative medicine applications.