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Objective: To investigate the orientation induced bone marrow mesenchymal stem cells (BMSCs) transplantation on spinal cord injury recovery of neurological function and appropriate time of transplantation. Method: 1) using adherent culture, purification BMSCs: Take the first three generations of BMSCs added with bFGF, EGF and B27 serum-free complex neural induction medium for induction, using immunocytochemistry to nestin identification of neural stem cells by adding serum-free culture , with its NF200 \\ GFAP identified neuronal differentiation, astrocyte-like cells. 3 days before cell transplantation to 5 Australia 2 deoxyuridine (Brdu) mark after induction of BMSCs; 2) using the Allen method produced models of spinal cord injury, 45 spinal cord injury model rats were randomly divided into three groups, each 15, A, B two groups were made in the model after one week, two weeks from the former two time points mark the site of injury Brdu transplant after induction BMSCs, C, control group, the original site of injury from injection of the same amount of physiological brine. Rats in each group after cell transplantation BBB score for motor function to determine recovery of neurological function and immunochemical staining fluorescence transplanted cell survival and differentiation (Brdu double labeled with NF-200) and the injury site regeneration of nerve fibers . HE staining area of spinal cord injury cavity size. Results: 1) in vitro amplification of BMSCs, the proliferation of fast expansion capability: the third generation BMSCs were highly homogeneous, showing monolayer cell bodies are mostly long spindle was swirling arrangement containing bFGF \\ EGF and B27 complex neural induction in serum-free medium can differentiate into neural stem cells, after serum was added to further differentiate into neuron-like cells and astrocyte-like cells, the corresponding expressing neurons, astrocytes markers NF200 and GFAP; 2) SCI cell transplantation after 1 to 6 weeks, A, B, C three groups of animals hindlimb neuromotor functions in varying degrees of recovery, A, B BBB scores in both groups were significantly higher than in group C ( P lt; 0.05). A, B groups immunofluorescence revealed that a large number of double-labeled Brdu and Nestin positive cells and some Brdu and NF200 double labeled positive cells filled in the area of spinal cord injury and damage to the host portion proximal and distal closely connected, see also the obvious nerve fiber regeneration, syringomyelia area was significantly smaller than in group C (P lt; 0.05). In group A, the change is more significant. Conclusion: In this study orientation induced bone marrow mesenchymal stem cells have the biological function of neural stem cells, and has further differentiation; transplanted BMSCs induced orientation spinal cord injury in rats can effectively promote recovery of neurological function, which, a week after transplantation than two weeks after the transplant.
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