The Effects Of Simple Or Combinate Transplantation With Different Stem Cells And Supporting Cells On The Neural Survival In Rats Subjected To Spinal Cord Transection

[Objective]In order to supply the experimental evidences for the effects of simple or cotransplant transplantation with nerve stem cells,olfactory ensheathing cell,haemopoietic stem cell and schwann cell on theⅧandⅨlayers neural survival in rats subjected to spinal cord transection.[Methods]The animals were divided into sham group(SG),control group(CG), simple transplantation of neural stem cells group(NSC),simple transplantation of olfactory ensheathing cells group(OEC),combination transplantation of neural stem cells and olfactory ensheathing cells group(NSC+OEC),simple transplantation of haemopoietic stem cells group(HSC),simple transplantation of schwann cells group(SC),combination transplantation of haemopoietic stem cells and schwann cells group(HSC+SC).Nerve stem cell,olfactory ensheathing cell,haemopoietic stem cell and schwann cell,were transplanted into the rostral,scar and caudal site of the transected cord of rats in the chronic phase(14 days post operation)by using the stereotaxic apparatus.6 months later,the spinal cord was obtained and prepared for paraffin section.Immunohistochemistry and hematoxylin stain were performed to explore theⅧandⅨlayers neural survival and host neural regeneration,further count the number and area of survival neuron.SPSS 11.5 statistical analysis software package was used to analyze the data.[Results]1.Positive staining of nestin,P75,CD34 and S 100 was detected respectively in the cultural cells by immunocytochemistry.2.To compare the quantity of theⅧneuron with every group,there were decreased significantly in control group compared with that of sham group.After simply transplantation with OEC or SC and combinate transplantation of NSC and OEC or HSC and SC,the quantity of neurons increased significantly in rats compared with that of control group,the quantity of neurons increased significantly in combinate transplantation of HSC and SC compared with that of simply transplantation with HSC or SC group (p＜0.05).To compare the quantity of theⅨneuron with every group,there were decreased significantly in control group compared with that of sham group.After simply transplantation with NSC,OEC or SC and combinate transplantation of NSC and OEC or HSC and SC,the quantity of neurons increased significantly in rats compared with that of control group,the quantity of neurons increased significantly in combinate transplantation of HSC and SC compared with that of simply transplantation with HSC or SC group(p＜0.05).3.To compare the area of theⅧneuron with every group,there were decreased significantly in control group compared with that of sham group.There were no detective changes of neural body after simply transplantation of NSC,OEC,HSC or SC,compared with that of control group.The body of neurons increased significantly in combinate transplantation of HSC and SC compared with that of simply transplantation with HSC or SC group(p＜0.05).To compare the area of theⅨneuron with every group, there were decreased significantly in control group compared with that of sham group.After simply transplantation with NSC,OEC or SC and combinate transplantation of NSC and OEC or HSC and SC,the area of neurons increased significantly in rats compared with that of control group.The area of neurons increased significantly in combinate transplantation of NSC and OEC or HSC and SC compared with that of simply transplantation with NSC,OEC,HSC or SC group. (p＜0.05).4.To observe the regeneration of serotonin axons,it grow into scar 2201am in cotrol group,2460μm in NSC group,560μm in OEC group,1820μm in HSC group,1770μm in SC group,890μm in NSC+OEC group,3365μm in HSC+SC group.To observe the regeneration of calcitonin gene-related peptide axons,it grow into scar 600μm in cotrol group,940μm in NSC group,460μm in OEC group, 680μm in HSC group,2090μm in SC group,3700μm in HSC+SC group.It was 130μm far from scar in NSC+OEC group.【Conclusion】1.The neurons inⅧandⅨlayers of host were atrophied or even died after spinal cord transection.2.There were more servival neurons in theⅧandⅨlayers of host following transplantation with OEC or SC simply and combinate transplantation of NSC and OEC or HSC and SC,only more servival neurons in theⅨlayers of host following transplantation with NSC simply.3.It was effective to prevent neurons from atrophia inⅨlayer but notⅧlayer after simple transplantation with NSC,OEC,HSC or SC following spinal cord transection.There were effective to prevent neurons from atrophia inⅧlayer using HSC and SC transplant combinatly contrasted to HSC or SC transplant simply.Contrasted to NSC or OEC transplant simply and HSC or SC transplant simply,It was a more effective method to preventⅨlayer neuron from atrophia using NSC and OEC transplant combinatly or HSC and SC transplant combinatly after spinal cord transection.4. Simple or combinate transplantation with NSC,OEC,HSC and SC was effective to promote serotonin and calcitonin gene-related peptide axonal regeneration after spinal cord transection.