Study Of Transplantion Of Human Marrow Stromal Cells After Orientiatedly Induced Into Neuron-like Cells

ObjectiveTo investigate the survival and differentiation of human marrow stromal cells transplanted into rat brain after induced into neuron-like cells. Methods1. Human marrow stromal cells (hMSCs) were isolated from human bone marrow by density gradient centrifugation method using lymphocyte isolation liquid.Cells were purified and expanded by plastic adherent culture.2. The third generation cultured hMSCs were induced with sodium ferulate for 24 hours, after pre-induced by bFGF and marked in cytoblast by Hoechst33342.3. The right middle cerebral artery occlusion (MCAO) model of SD rats was produced by obstructing MCA using a single strand nylon thread, and we reperfused the artery after 2 hours.4. The induced and marked hMSCs about 5 × 10~6/0.8ml were transplanted into the right internal carotid artery of MCAO rats after reperfusion 24 hours.5. Two weeks after the hMSCs transplantion, we opened the thoracic cavity, and washed arteriovenous through heart and fixed the braing by 4% paraformaldehyde, and the rat brain was anhydrated by 30% sucrose.6. Frozen sections of rat brain near the optic chiasma were made paralleling the coronal plane. The expression of MAB1281, nestin and NSE in the brain tissue were detected by immunofluorescence staining. The cells marked with Hoechst were observed under the fluorescence microscope.Result...