Objective: Adipose-derived mesenchymal stem cells (AMSCs), as adult stem cells, have potential to differentiate into various lineages such as cartilage. In order to use equine AMSCs in treatment of diseases including musculoskeletal disorders, it is obligatory to understand the effect of donor age on the differentiation potential of these cells. Material and Methods: To isolate AMSCs, fat samples were collected from three mares aged 3, 6 and 10 years old and were transported to the laboratory. Immediately, cells were isolated by mechanical and enzymatic (Collagenase I) and were cultured in optimized conditions. The cultured cells were passaged until passage 3. To assess the chondrogenic potential of passage 3 cells, they were differentiated under the chondrogenic medium supplemented with TGF-β3 and BMP-6 for 21 days. High-density cell culture system (micromass) was chosen. Chondrogenic differentiation was confirmed by toluidine blue staining and Pellet sizes were measured by microscope. Result and Conclusion: All cultured cell pellets were differentiated to chondrocytes under chondrogenic differentiation condition when compared to control group. Differences in the chondrogenic differentiation capacity of equine AMSCs harvested from donor animals of different age were observed. Chondrogenic potential decreased with age. After 21 days, Cell micromasses from younger donors (3 and 6 years old horses) were larger than 9 years old horse. In conclusion, increasing age reduced chodrogenic differentiation potential of AMSCs.

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