(W1253) CT PROMOTES SPHEROID FORMATION AND EXTRACELLULAR VESICLE SECRETION OF HUMAN UMBILICAL CORD MESENCHYMAL STEM CELLS AND ITS ROLE IN SKIN INJURY TREATMENT
Objective: Human umbilical cord mesenchymal stem cells (huc-MSCs) are a promising source for therapeutic applications due to their pluripotency. Extracellular vesicles (hucMSC-EVs) secreted by huc-MSCs contain bioactive molecules and have shown therapeutic potential. However, conventional 2D cultures face challenges like limited space, inconsistent cell quality, and inefficient use of culture area. Scaffold-free MSC spheroid cultures offer a more in vivo-like environment, preserving cell characteristics and optimizing space. Despite these advantages, spheroid culture faces challenges in spheroid formation and EV yield. This study explores how combining Chroman 1 and Trans-ISRIB (CT) enhances MSC spheroid formation, EV secretion, and their therapeutic potential for skin injury.
Methods: The CT combination was screened for its ability to promote MSC spheroid formation and EV secretion using CEPT. Transcriptome sequencing and PCR were used to explore the mechanisms behind CT’s effects on spheroid formation. MSC EV-related gene expression was analyzed, and EV yield was quantified by nanoparticle tracking analysis (NTA). The therapeutic effects of CT-3D MSC-EVs on skin injury were assessed in vitro using HUVECs and fibroblasts, and in vivo in a mouse skin injury model.
Results: CT significantly promoted MSC proliferation and increased paracrine factor expression. In spheroid cultures, CT enhanced spheroid formation and upregulated EV-related (CD9, CD81, CD63, Alix) and stemness (Nanog, Sox2, Oct4) genes. Transcriptome analysis and PCR revealed that CT promoted spheroid formation by upregulating cell adhesion molecules. Scratch assays demonstrated that CT-3D MSC-EVs enhanced migration of HUVECs and fibroblasts, outperforming 2D-MSC-EVs. Tube formation assays confirmed that CT-3D MSC-EVs better promoted HUVEC tube formation. In vivo, CT-3D MSC-EVs accelerated skin wound healing and dermal collagen regeneration in mice.
Conclusion: The CT combination enhances MSC spheroid formation and EV secretion by upregulating cell adhesion molecules, improving the therapeutic effects of spheroid-derived MSC-EVs for skin injury. These findings highlight the clinical potential of MSC-EVs in therapeutic applications
