Species-specific optimisation of cryopreservation media for goat and buffalo adipose-derived mesenchymal stem cells

Adipose-derived mesenchymal stem cells (ADSCs) are promising for clinical and veterinary applications due to their ease of isolation, high yield, and multilineage differentiation potential. Effective cryopreservation is vital to ensure their availability for large-scale applications. This study evaluated cryopreservation strategies for goat (gADSCs) and buffalo (bADSCs) ADSCs, using combinations of intracellular (dimethyl sulfoxide, DMSO) and exocellular cryoprotectants, including fetal bovine serum (FBS), polyethylene glycol (PEG), trehalose, bovine serum albumin (BSA), and dextran. Post-thaw parameters such as viability, recovery, metabolic activity, clonogenicity, oxidative stress, apoptosis, and senescence were assessed. Results revealed species-specific differences in cryopreservation requirements. gADSCs were optimally preserved in a medium with 5 % DMSO, 3 % FBS, 2 % PEG, 3 % trehalose, and 2 % BSA, while bADSCs performed best in an FBS-free medium containing 5 % DMSO, 2 % PEG, 3 % trehalose, and 2 % BSA. DMSO-FBS formulations supported high recovery and metabolic activity but were associated with increased oxidative stress and apoptosis. Dextran-based cryomedia effectively preserved gADSCs but failed to maintain bADSC functionality. Biochemical composition analysis indicated significantly higher lipid content in bADSCs, likely influencing cryopreservation efficacy. These findings underscore the need for tailored cryopreservation strategies to address species-specific differences. Incorporating exocellular cryoprotectants reduced FBS dependency, minimised oxidative damage, and maintained functional attributes. This study highlights the potential for optimised, cost-effective biobanking solutions that accommodate species-specific requirements, advancing the use of ADSCs in veterinary and translational research.