Regenerative medicine aims to replace or repair damaged organs and tissues including the injured joints and tendons. Research on the use of mesenchymal stem cells (MSCs) in tissue regeneration has generally been driven by the needs of human and veterinary medicine. MSCs are one of the best candidates in treatment of musculoskeletal disorders. Nonetheless, the most suitable condition for their chondrogenic differentiation is under investigation. In this work, about 2-3 grams of equine fat tissue was isolated and transported to the laboratory. MSCs were isolated using mechanical and enzymatic digestion. Isolated cells were cultured in optimum medium until passage 3 (P3). To analyze chondrogenic differentiation, the P3 cells were cultured under tree different conditions: 1) basic medium as control, 2) differentiation medium without TGF-β3 and 3) differentiation medium supplemented with 10 ng/ml TGF-β3. Chondrogenic differentiation was applied for 21 days in 3D micromass culture system and medium was changed every 4 days. Isolated MSCs were plastic-adherent, fibroblast-like and expressed specific markers. After the period of chondrogenic culture, no significant differences were observed in pellet sizes among 3 groups. Histological results confirmed chondrogenic differentiation in groups containing chondrogenic medium (2 and 3) compared with control group. More coherent extracellular matrix and typical lacuna formation confirmed better chondrogenic differentiation in the group supplemented with TGF-β3. The results suggest that addition of TGF-β3 to the standard chondrogenic medium is not required for chondrogenic differentiation of equine adipose-derived MSCs, but it promotes the differentiation.

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