Experimental Study on Isolation,Culture and Identify of Rabbit Bone Marrow-derived Mesenchymal Stem Cells in Vitro

Objective To explore the approaches of isolation,culture and identify the rabbit bone marrow-derived mesenchymal stem cells in vitro.Methods MSCs were extracted from rabbit bone marrow(5.0 ml) and purified via Density gradient centrifugation,then proliferated in vitro.The cell growth was observed under Phase-contrast microscope and the growth curve of cell proliferation was drawn accordingly.Light microscopy were used to study the morphologic features and indirect immunofluorescence was used to examine the expression of cell surface antibody of CD44 and CD34.Results MSCs of rabbit were isolated,cultured in vitro.The proliferation of MSCs decreased while the cell proliferated more generations.The MSCs of 3rd and 5th generation proliferated more rapidly than those of 7th and 9th generation.These cultured cells showed immunoreactivity to CD44 but not CD34.Conclusion Purified MSCs of rabbit could be obtained via Density gradient centrifugation.Cells of 3rd,5th generation with high proliferation were effective to the further experiment.