Enhancing the clinical potential: Quality assessment of adipose tissue-derived Mesenchymal Stem/Stromal Cells for therapeutic use.

[PURPOSE] Despite ongoing debate surrounding mesenchymal stem/stromal cells (MSCs), the number of clinical trials involving MSCs has continued to increase due to their strong therapeutic potential. These cells exhibit immunosuppressive and neuroprotective properties, making them a promising approach for treating neurodegenerative diseases. Given the growing interest in stem cell-based therapies, it is essential to establish specific protocols for manufacturing adipose-derived MSCs. This study outlines the process of obtaining an ATMP cell product from ASCs, evaluating cell quality, isolation efficiency, and optimal conditions for cryopreservation, preparation, and transport.

[MATERIALS AND METHODS] Cells were isolated from fresh, cryopreserved, and 4°C-stored adipose tissue to evaluate how different storage conditions affect cell quality. In addition, cell viability in different transport solutions over time was examined. To evaluate the therapeutic potential of ASCs, cells were cultured in cerebrospinal fluid (CSF) and neural gene and protein expression, as well as the secretome profile, were analysed.

[RESULTS] Our findings show that isolating ASCs immediately after liposuction yields the highest viable cell count. Adipose tissue can be stored at 4 ​°C for up to 48 ​h without affecting cell population doubling time, but cryopreservation negatively impacts ASCs' clonogenicity. For optimal long-term preservation, a medium with 5 ​% platelet lysate and 10 ​% DMSO is recommended. Additionally, we found that 0.9 ​% NaCl is the best transport medium for maintaining cell viability. CSF exposure induced neural markers (NG2, A2B5, Nestin, β-III-tubulin, S-100-β) and morphological changes, suggesting neural-like differentiation.

[CONCLUSIONS] These findings contribute to the development of protocols for MSC therapy in neurodegenerative diseases.