This study was designed to evaluate the in vitro chondrogenic differentiation potential of equine adipose tissue-derived mesenchymal stem cells (AT-MSCs) in response to different culture conditions. | Turkish Journal of Biology Turk J Biol (2016) 40: 360-368 © TÜBİTAK doi: Research Article Enhancement of chondrogenic differentiation potential of equine adipose tissue-derived mesenchymal stem cells using TGF-β3 and BMP-6 1 1,2, 1,2 Milad SHADEMAN , Abbas PARHAM *, Hesam DEHGHANI Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran 2 Embryonic and Stem Cell Biology and Biotechnology Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran 1 Received: Accepted/Published Online: Final Version: Abstract: This study was designed to evaluate the in vitro chondrogenic differentiation potential of equine adipose tissue-derived mesenchymal stem cells (AT-MSCs) in response to different culture conditions. Fat tissue cells after collection were cultured in optimized conditions until passage 3 (P3). P3 cells were cultured as micropellets under different chondrogenic conditions for 21 days in 5 groups: basic medium as a control, basic chondrogenic medium (BCM), BCM supplemented with transforming growth factor beta 3 (TGF-β3; 10 ng/mL), BCM supplemented with bone morphogenetic protein 6 (BMP-6; 10 ng/mL), and BCM supplemented with both TGF-β3 and BMP-6. The growth rate of pellets was measured and differentiation progression was assessed by specific stainings and gene expression profiling. Results showed that the largest pellets of cells belonged to the group containing both growth factors. All treated cells showed different levels of chondrogenic differentiation compared to the control group (P ) greater than the group with only TGF-β3 (Table 2). Table 1. Nucleotide sequences of the primer sets used for RT-PCR. Genes GenBank accession number Primer pairs Annealing temperature (°C) Amplicon size (bp) Equine GAPDH NM_001163856 F: TGTCATCAACGGAAAGGC R: .
