Kandpal, ManjulVerma, Rakesh Kumar2019-02-022019-02-022018-07http://krishikosh.egranth.ac.in/handle/1/5810093314The adult stem cell therapy is a blooming area of clinical research and relevance. The therapeutic benefits of mesenchymal stem cells in veterinary regenerative medicine are numerous as they are in human medicine. The mesenchymal stem cells in canines can be potentially used for cell based therapy to regenerate damaged or lost neuronal cells. The leitmotif of this study was differentiating cBM-MSCs into neurocytes in vitro. A healthy nondescript male dog of 3 years of age and 24kg weight was used for this study. The bone marrow aspirate was aseptically collected from ileac crest of the dog after administration of general anaesthesia by 2.5% thiopentone sodium till effect and atropine sulphate and diazepam as pre-anaesthetics. The bone marrow aspirate was processed in laboratory within 4 hours of its collection. Mesenchymal stem cells were isolated by density gradient centrifugation technique and cells were seeded in T-25 tissue culture flasks. After attainment of 80-90% confluence, first passage was performed by using Trypin-EDTA to expand the cell population. Cells were passaged till 4 passages and then grown in neurogenic differentiation media for 21 days in CO2 incubator at 37°C and 5% CO2. The morphology of cells was transitioned from spindle elongated shape to ring or net like shape. The cells were characterized at regular intervals by staining with the Nissl body stain after 21 days with positive result which coloured the nuclei blue-violet. The RT-PCR for the genes NSE and NFM revealed bands on days 21 confirming the differentiation process. The sample on gelatin scaffold and on pellet when subjected to scanning electron microscopy revealed net shape cells with cell aggregates. On the basis of this study it was concluded that MSCs possess potential to transdifferentiate to neurocytes when cultured in neurogenic media in vitro and the identification techniques are successful. The meticulous exploitation of MSCs in tissue engineering and regenerative therapy in veterinary medicine is the need of the hour.ennullThesis