Optimised cryopreservation preserves functional competence of goat adipose-derived mesenchymal stem cells and is associated with stress-adapted mitochondrial and paracrine features

Cryopreservation is essential for mesenchymal stem cell banking, but its effects on adipose-derived stem cell (ADSC) function remain debated. This study evaluated an optimised cryopreservation protocol for goat ADSCs using phenotypic, functional, mitochondrial-associated, conditioned-medium, proteomic, and wound-healing assays. Cryopreserved ADSCs retained morphology, viability, clonogenicity, proliferation, adherence, immunophenotype, and trilineage differentiation, comparable to those of fresh ADSCs. Post-thaw cells showed increased MitoTracker Green signal, JC-1 red/green ratio, ATP content, relative mtDNA abundance, and SOD2-associated immunofluorescence, together with lower H ₂ O ₂ -induced ROS- and autophagy-associated fluorescence. Conditioned-medium proteomics suggested exploratory shifts toward extracellular matrix-, anti-protease-, and redox-associated proteins. Functionally, cryopreserved ADSCs maintained inflammatory cue-directed migration, modulated macrophage surface marker expression, and reduced oxidative stress-associated injury readouts in goat dermal fibroblasts. In a goat excisional wound model, ADSC treatment improved wound-closure kinetics and supported histological repair. Overall, optimised cryopreservation preserved the functional competence of goat ADSCs and was associated with stress-adapted post-thaw features.