Int.J.Curr.Microbiol.App.Sci (2020) 9(6): 1380-1387 1380 Original Research Article https://doi.org/10.20546/ijcmas.2020.906.170 Assessment of Invitro Differentiation Potential of Canine Bone Marrow Derived Mesenchymal Stem Cells to Skeletal Myocytes R. S. Sandhu 1* , N. S. Jadon 1 , P. Pandey 2 , D. Bodh 1 , P. Joshi 1 , M. Kandpal 1 , R. Saini 1 and A. Kumar 1 1 Deptartmen of Veterinary Surgery & Radiology, 2 Department of MBGE, CBSH, G.B.P.U.A. and T., Pantnagar (U.S.Nagar), Uttarakhand (Pin: 263145), India *Corresponding author ABSTRACT Introduction Mesenchymal stem cells (MSCs) or stromal progenitor cells is a population of stem cells. Mesenchymal stem cells regenerate tissues of mesenchymal lineages by differentiation, and trans-differentiate into cells derived from other embryonic layers, including neurons and epithelia in skin, lung, liver, intestine, kidney, and spleen. These cells form an elemental part of haemopoeitic micro environment which supports blood cell development. These cells are multipotent in nature as they can differentiate into some mesodermal lineages including adipocytes, osteocytes and chondrocytes . Additionally MSCs have a distinct artifice to suppress immuno mediated inflammatory responses (Griffin et al., 2013). Mesenchymal stem cells which are isolated from bone marrow have exhibited to infer a skeletal myogenic outcome (Mizuno, 2010). International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 9 Number 6 (2020) Journal homepage: http://www.ijcmas.com Mesenchymal stem cells are multipotent/pluripotent stromal cells that can differentiate into a diversity of cell categories depending upon the micro-environment to that they are exposed. The multipotent differentiation potential of MSCs is generally towards a constrained sum of cell types familiar to the tissue of origin and maintains the capacity for self-renewal. To assess the differentiation potential of MSCs to skeletal myocytes a male mongrel dog of 3 years was used to carry out the research. The marrow derived mesenchymal stem cells were harvested with mononuclear cells and cultivated in monolayer culture in T-25 tissue culture flasks till they attained of 80-90 % confluency. At 4 th passage the cells were driven towards skeletal myogenesis by addition of skeletal myocytes differentiation medium containing 5-azacytidine, foetal horse serum, basic fibroblast growth factor, dexamethasone and hydrocortisone. The cells which differentiated showed positive results of RT-PCR for the early myogenic differentiation factors Myf5 and Myogenin and Hoechst 33342 staining. The Hoechst 33342 stainin, RT- PCR analysis and scanning electron microscopy leads to the positive result for differentiated skeletal myocytes in vitro. The differentiated skeletal myocytes can be used in tissue engineering and veterinary therapeutics for reparing the damaged skeletal myocytes. Keywords MSCs, Skeletal myocytes, Myf5, Myogenin, Hoechst 33342. Accepted: 18 May 2020 Available Online: 10 June 2020 Article Info