ISOLATION, CHARACTERIZATION, AND DIFFERENTIATION OF MESENCHYMAL STEM CELL FROM RAT BONE MARROW IN VITRO

Author(s): Ranjith Santhosh Kumar DS, Senthil Kumar P*, Surendran L, Sudhagar B and Vinodhini J

Abstract: Bone marrow mesenchymal cells have been identified as a source of pluripotent stem cells with varying degrees of plasticity in humans. MSCs have the potential to differentiate into different cell types such as osteoblast, chondroblast, adipoblast. This research describes the pioneering experiment of isolation and differentiation rat bone marrow stromal cells into the osteoblast and adipoblast cells lineages. Bone marrow stromal cells were isolated from long bones of rat, based on the adherent properties of the MSC. The stromal cells obtained by direct plastic adherence were characterized by fluorescent activating cell sorting (FACS) for established hematopoietic and non-hematopoietic markers. Under the determinate effect of culture conditions, MSCs were differentiate done osteogenic and adipogenic cell lines detected by Alizarin red and oil-red O staining invitro. Bone marrow samples from rats yielded 4-5 million bone marrow mononuclear cells/ml per femur. The MSCs culture is being investigated with microscopic observations. They form elongated or spindle-shaped fibroblast-like cells. Cell sorting by the surface marker expression is used in the current studies for confirmation of rat MSCs in the culture. Moreover, the cells differentiated toward osteoblasts and adipocytes were characterized morphologically by Alizarin Red staining and Oil Red O staining. This research consigns new ideas to develop noble markers and play a vital role in tissue engineering.

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