APPROACHES TO ISOLATE,CULTURE AND IDENTIFY THE BONE MARROW-DERIVED MESENCHYMAL STEM CELLS OF RABBITS

Objective To explore the approaches to isolate,culture and identify the bone marrow-derived mesenchymal stem cells(BM-MSCs) of rabbits. Methods BM-MSCs were isolated from the bone marrow(5.0 mL) of the newborn rabbit via density gradient centrifugation.Light and electron microscopy were used to study the morphologic features and immunocytochemistry to examine the expression of cell surface antigens.The growth curve of cell proliferation was obtained based on the observation of the proliferation status of 1st,3rd,5th,8th,and 10th generation. Results BM-MSCs of 1st,3rd,5th generation were very purified and all in the whirlpool-shape.They proliferated more rapidly than those of 8th and 10th generation.These cultured cells showed immunoreactivity to CD44 and CD90 but not CD34 and low-differentiation under the electron microscopy. Conclusion(Purified BM-MSCs) can be obtained via this protocol.Cells of 1st,3rd,5th generation with high proliferation are fit to the further experiment.