(F1231) Induced Pluripotent Stem Cell-Derived Mesenchymal Stromal Cells: A Scalable and Versatile Source of Extracellular Vesicles for Regenerative Medicine
Center for iPS Cell Research and Application (CiRA), Kyoto University Kyoto, United States
Abstract: Induced pluripotent stem cell-derived mesenchymal stromal cells (iMSCs) represent an emerging approach for the production of functional extracellular vesicles (EVs) for regenerative medicine. Here, we emphasize the advantages of iMSCs as a reliable, highly expandable cell source of EVs (iEVs) and a platform for engineering EV cargo. These characteristics make iMSCs a promising alternative to tissue-derived mesenchymal stromal cells (MSCs), which are subject to donor variability. iEVs and EVs isolated from MSCs were characterized using transmission electron microscopy (TEM), flow cytometry, and interferometry to assess size, concentration, and morphology. iMSCs exhibited a 3X higher yield of iEVs, along with increased expression of exosome markers compared to MSCs. Proteomic analysis of EVs and iEVs provided valuable insights into their therapeutic cargo, including key molecules involved in immunomodulation and tissue repair. Additionally, glycan analysis identified distinct glycan signatures that play a critical role in cell recognition, adhesion, and internalization, important processes for the delivery of therapeutic cargo. Comparative functional assays conducted on fibroblasts, macrophages, and human umbilical vein endothelial cells (HUVECs) revealed that both iEVs and EVs supported key cellular processes essential for tissue repair. Notably, iEVs demonstrated greater consistency in their effects, particularly in their ability to modulate macrophage polarization from a pro-inflammatory M1 to an anti-inflammatory M2 phenotype. In vivo studies using a murine wound model showed that iEVs effectively reduced inflammation, promoted M1-to-M2 macrophage polarization, and accelerated wound healing. Furthermore, engineering approaches, such as genetic modification and preconditioning of iMSCs, further enhanced the immunomodulatory properties of iEVs. These findings support the use of iMSCs as a versatile and reliable option for EV-based therapies, offering a scalable and reproducible solution for regenerative medicine. Additionally, EV engineering provides an opportunity to extend their therapeutic potential to address a range of unmet medical needs.
