Human Umbilical Cord Mesenchymal Stem Cells Transplantation For Treatment Of Cerebral Hemorrhage Injury In Rats

OBJECTIVE:To study the effects of bone marrow mesenchymal stem cells (BMSCs) on recovery of neurological function in cerebral hemorrhage disease mouse modle,and to explore the therapeutical and potential mechanism.METHODS:We can separate green fluorescent protein transgenic rat BMSCs by density gradient centrifugation and adherence screening method.we can inject collagenase to animal models right globus pallidus to induce cerebral hemorrhage.the adult male sd rats animals were randomly divided into saline group,model group and transplantation group. right globus pallidus injected by2ul collagenase which was diluted with sterile saline.saline group injected with normal saline.transplantation group were injected with BMSCs by the same method after2h.saline group and model group were not treated.we can examine the the recovery of rat nerevous system function with mNSS, observe the behavioral changes in the morris water maze and the changes of rat brain tissue by MRI.rat were killed on ld,3d,7d,14d and28d.we can observe the BMSCs survival and the changes of inflammatory factors,neurotrophic factors and other indicators in the brain.RESULTS:By the way of density centrifugation combine with screening method to separate bone marrow mesenchymal stem cells, subsequently transfer of culture and amplification and differentiate in vitro. The separated cells was detected by flow cytometer, result display the cells express CD90and CD29, accord with the characteristics of BMSCs,such as the form and cell-surface marker.The cells possess favourable self-renewal ability and multi-directional differentiation potential, confirm the cells we separated is BMSCs.2.Model was incuced by injecting collagenase into animal models right globus pallidus.We can observe the brains by MRI after six hours.T1WI sequence shows the corresponding level of the right striatum small piecesdensity,suggest the model was successful.3.After BMSCs transplantation we found that transplantation group mNSS score is lower than model group.4.The expression and apoptosis of the transplantation group significantly reduced,but the expression of BDNF protein in the brain hemorrhage significantly increased.Brain tissue can be seen a rery small amout of GFP+NeuN,GFP+GFAP positive cells. CONCLUSION:Bone marrow mesenchymal stem cells could separated and amplification in vitro. model incuced by injecting intoanimal models right globus pallidus,this method can be used for the treatment of hematencephalon in rats.The BMSCs transplantation can significantly reduce TNF-a and apoptosis and improve the expression of BDNF protein in the brain hemorrhage.Bone marrow mesenchymal stem cells can differentiated into neuron-like and glial cell-like cells after transplantation, have effect of improving spacial learning-dysmnesia of rats. Transplantation of BMSCs is safety and effective, suggesting it can be a new way.