Evaluation of Batch-To-Batch Variability and Efficacy Of IPSC-Derived Mesenchymal Stromal Derived Extracellular Vesicles in Mitigating Osteoarthritis Associated Inflammation In Vitro

Abstract Background Mesenchymal stromal cell-derived extracellular vesicles (MSC-EVs) hold promise as a cell-free therapy for various diseases, including osteoarthritis (OA), due to their immunomodulatory and anti-inflammatory properties. However, the need for large-scale expansion of MSCs to obtain MSC-EVs for clinical use often leads to senescence-related changes and loss of stem-like properties. In this scenario, induced pluripotent stem cell (iPSC)-derived MSCs (iMSCs) offer the unique opportunity to address obstacles associated with traditional MSC-based therapies. Methods In this study, a xeno-free (XFS) medium was used for long-term expansion of both MSCs and iMSCs, and subsequent isolation of MSC- and iMSC-derived EVs. Characterization of both cells and EVs was conducted across different passages, and the anti-inflammatory potential of EVs and iEVs was assessed using an in vitro model of osteoarthritis. Results Long-term expansion of MSCs resulted in cellular senescence and a reduction in trilineage differentiation capacity by passage five, accompanied by diminished anti-inflammatory properties of EVs. On the other hand, iMSCs exhibited batch-to-batch variability in differentiation potential and the biological properties of iEVs. However, the effects of iMSC-EVs are prolonged compared to MSC-EVs, providing a prolonged window of activity for therapeutic purposes. Conclusions While iMSC-EVs demonstrated prolonged effects compared to MSC-EVs, heterogeneity among iMSC batches present significant challenges in utilizing iMSC-derived EVs as a reliable tool for clinical OA treatment. Addressing these limitations is crucial for advancing the translational potential of iPSC-derived therapies in the management of osteoarthritis and other diseases.