Experimental Methods Study of the Investigation on Inducement and Differentiation of Rat Whole Bone Marrow Cells into Nerve Cells in vitro

Objective: To find a method of differentiating of the whole bone marrow cells into nerve cells and observe their developing way on morphology. Methods:The whole bone marrow cells were cultured after aspirating from the femur and tibia of albino Sprague-Dawley adult rats(Harlan-Sprague-Dawley) and taking off the erythrocytes by hypotonic method. The morphology of the cultured cells in different stages was observed by microscope and the expression tendency of nestin and neuronal nuclei(NeuN)positive cells ere from bone marrow cells to nerve cells was tracked by fluoresce in activated cell sortor(FACS). In addition, these cells were identified by immunocytochemistry staining. Results:Bone marrow cells began to differentiate at the tenth day after culture and gradually developed to become neuron-like cells. Expression rate of nestin positive cells peaked to 22.7% at the fifteenth day and NeuN's peak at the thirtieth day which rate of 41.2%. Whereas, the percentage of nestin positive cells was 5.9% at the time. The nerve stem cells, neurons and glial cells were identified by immunocytochemistry. Conclusion:The experiment was successed in finding a method to make the whole bone marrow cells differentiation into nerve cells, which possesses simple, fast and highly effective.