Abstract: Mesenchymal stem cells (MSCs) are highly recognized in regenerative medicine due to their multipotent differentiation and immunomodulatory properties. Recent studies suggest that the immunomodulatory effect of MSCs is mainly mediated through paracrine factors, with exosomes being critical players. This study focuses on the immunomodulatory effects of MSCs-derived exosomes. MSCs were isolated from bone marrow and cultured in Cell Vive™ MSC serum-free, xeno-free media, GMP. Exosomes were isolated from the conditioned medium using a tetraspanin-based MojoSort ™ human microbead exosome isolation kit, with purity confirmed via flow cytometry, western blot, and nanoparticle tracking analysis. The cargo of MSCs-derived exosomes was characterized using LEGENDplexTM assay, a multiplex flow cytometry-based immunoassay system, revealing a diverse array of bioactive molecules crucial for tissue repair and immunomodulation. We investigated the immunomodulatory effects of these exosomes on CD3/CD28 activated T cells. Exosomes were depleted from the MSCs conditioned media, and T cells were cultured in conditioned media (CM) with and without exosome-depletion. Data demonstrates that exosomes-depleted MSCs CM exhibits reduced modulation potency in suppressing activated T cell proliferation, and cytokine profiles from MSCs-derived exosomes were analyzed to highlight potential cyotokine candidates contribute the MSCs immunomodulation capabilities. These findings highlight the potential of MSCs-derived exosomes as a cell-free therapeutic option in regenerative medicine.
