Study On Treatment Of Autologus Activated Schwann Cells And Human Embryonic Neural Stem Cells In Spinal Cord Injury

Objective: To observe the effect of acute spinal cord injury in Wistar rats by co-transplantation of Schwann cells and human neural stem cells. Methods: Autologus activated Schwann cells from the femoral nerve of the mature Wistar rats were isolated and cultured by primary cell culture and purified by rapid trysinization and different velocity of adhesion to the bottom of culturing bottle. A neural stem cell line which had ability to form clones was isolated from the cortex of human embryonic brain by serum-free cell culture. The clone was passaged continuously for 4 generations in order to acquire a large amount of clones, and indirect immunofluorescence was used to detected Nestin antigen. The two type of cells were co-cultured and analysed the compatibility between them. Then the 40 adult Wistar rats, weight 200±30g, were divided into 4 groups: Blank group, SCs groups, NSCs group and Compound Cells group and all rats were compressed on T10 (Allen's striking way) for about 3min by 15g. The grafts of AASCs and/or NSCs or none were transplanted to the site of spinal cord injury by microinjection.Then we observed their recovery of the lower extremities with BBB score and SEP&MEP.After 15 weeks, two rats of each group selected in random were injected 10%BDA in the double cortex of brain for anterograde labeling the injuried spinal cord . After two weeks, these rats were killed. The injuried spinal cord including the injuried segment were taken out and cut into slim for 5μm thick at - 19℃.The slims were stained by cy3, HE, and immunohistochemistry was used to detect NF. The image was analysed by Image pro plus and the results were analysed by ANOVA and SNK-q.Results: NSCs were positive for Nestin. SCs were defined by appearance under microscope.The two type of cells have good compatibility. The recovery of sense and motion function in Compound Cells group and SCs group were better than the other two groups. The regenerated neurofibril stained by cy3, HE and NF could be seen in the injuried position in Compound Cells group and AASCs group, almost nothing in the other two groups. There was no apparent difference between Compound Cells and SCs group or NSCs and Blank group in BBB score(p>0.05). But not for the other situations(p<0.05).Conclusion: Transplantation of autologus activated Schwann cells and Human embryonic neural stem cells can guide the regeneration of neurofibril.