Neural Stem Cells Pretreated By BDNF Transplantation On Spinal Cord Injury

Chapter oneResearch Background: The neural stem cells with self-renewal,multilineage differentiation and self-proliferation can be isolated and cultured from the nerve tissue of mice.Research aim: The aim of this study was to isolated cells with self-renewal,proliferation,and differentiation of C57/BL-6 from mouse spinal cord neural stem cells in the plant,according to the observation and analysis of its biological properties,screened from active proliferation and could differentiate into neurons ability of neural stem cell line could be used as seed cells for subsequent research.Research method: The study by the monoclonal cells in serum-free culture technology,points out different neural stem cells from neonatal mouse spinal cord after passage third,by immunofluorescence on the specific markers of NESTIN were identified and cultured for seven days after induced by 10% fetal bovine serum,cell immunofluorescence on the differentiation of neuron specific markers of product Tuj1,GFAP of astrocytes were identified,screening 3 strains of stable passage ability of neural stem cells were tested for proliferation by CCK-8 method,and then filter out the strongest proliferative ability of neural stem cells in the later study.Result and Conclusion: Visible under the microscope isolated neural stem cells cultured for third days in the formation of neurospheres confirmed that the cells have the ability to self renew;spread to the third generation of neural stem cell specific marker NESTIN expression positive results,combined with morphological observation confirmed that the extracted cells for neural stem cells;differentiation of fetal bovine serum by 10% 10 days later,Tuj1 GFAP,immunofluorescence test positive results that neural stem cells can differentiate into neurons,astrocytes and neural stem cells have confirmed the differentiation of neural stem cells;3 strains of CCK-8 test results of selected strong proliferation of neural stem cells as seed cells for the next step.Chapter twoResearch Background: Neurotrophic factors play a neuroprotective role in promoting the proliferation of neural stem cells and inducing them to differentiate into neurons.Research aim: The purpose of this study was to evaluate the effects of brain-derived neurotrophic factor,ciliary neurotrophic factor and transforming growth factor(TGF)on proliferation,differentiation and apoptosis of neural stem cells in vitro.Research method: Using CCK-8 method to monitor the brain-derived neurotrophic factor,ciliary neurotrophic factor,nutrition factor,transforming growth factor on proliferation of neural stem cells function;to detect the expression of different neurotrophic factors on differentiation of neural stem cells were neuron specific markers of beta-Tubulin III by immunofluorescence and fluorescence microscope to count the differentiation of product neurons,and then filter out the strongest neurotrophic factor induced neuronal orientation;to observe the effects of different neurotrophic factors on apoptosis rate of neural stem DAPI staining method,screening neurotrophic factor inhibition of stem cell apoptosis process of optimal nerve nature.Result and Conclusion: Immunofluorescence results showed that cell screening positive expression of Nestin and further confirmed for the study cells into neural stem cells;CCK-8 assay showed that the proliferation of neural stem cells for BDNF treatment group obviously ciliary neurotrophic factor group(P<0.01)is greater than the transforming growth factor group(P<0.05)higher than the control group(P<0.01),that brain-derived neurotrophic factor has a larger advantage in promoting the proliferation of neural stem cell ability;neuron counting showed that transforming growth factor treatment group was significantly higher than that of BDNF group(P<0.05)is less than the ciliary neurotrophic factor(P=0.05)treatment group was significantly higher than the control group(P<0.01).It is shown that the transformation of neurotrophic factor in inducing differentiation of neural stem cells into neurons ability the most advantage;DAPI staining The monitoring results showed the apoptosis rate of BDNF treatment group was significantly lower than that of ciliary neurotrophic factor treatment group is less than the transforming growth factor group(P<0.05)was significantly higher than the control group(P<0.01),description of brain-derived neurotrophic factor in the optimal maintenance of neural stem cells in normal life effect.Chapter threeResearch Background: Neurotrophic factors play a role in the recovery of neurological function.Research aim: The aim of this study was to assess the in vivo environment observation of brain-derived neurotrophic factor,ciliary neurotrophic factor,transforming growth factor of animal models for BBB score of hind limb function and rotation balance test and experiment test,and the slope angle change of the neuron degeneration effect on spinal cord injury;and evaluate its therapeutic effect on animal models of spinal cord injury,and selected the treatment effect is the most obvious neurotrophic factor,which lays the theoretical foundation for follow-up study.Research method: The test score improvement different neurotrophic factors on mouse hindlimb function after spinal cord hemisection by hindlimb function BBB,screening of neurotrophic factor therapy is best;the rotary balance test and experimental test to evaluate the therapeutic effect of slope angle of different neurotrophic factors on mouse behavioral function after spinal cord hemisection,selected from improving neurotrophic factor has the strongest effect;to observe the effects of different neurotrophic factor on neurons injury of spinal cord after silver staining,were selected for follow-up study at least denatured neurons neurotrophic factor.Result and Conclusion: Hindlimb function BBB scores showed that BDNF treatment group was significantly higher than that of ciliary neurotrophic factor(P<0.01)treatment group was higher than that of transforming growth factor(P<0.05)group was significantly higher than that of sham operation group(P<0.01),that brain derived business in promoting the ability of God son hindlimb functional recovery after spinal cord injury is highest;rotation balance the test results showed that CNTF treatment group was higher than that of brain-derived neurotrophic factor(P<0.05)treatment group was higher than that of transforming growth factor(P<0.01)group was significantly higher than that of sham operation group(P<0.01),angle of slope test test results show that BDNF treatment group was significantly higher than that of transforming growth factor group(P<0.01)Gao Yujie ciliary neurotrophic factor(P<0.01)group was significantly higher than that of sham operation group(P<0.01),brain derived neurotrophic factor can effectively promote the spinal cord injury The effect of rehabilitation after silver staining;neuron counts showed degeneration of ciliary neurotrophic factor in treatment group was significantly lower than that of transforming growth factor(P<0.05)group was lower than that of brain-derived neurotrophic factor(P<0.05)group was significantly lower than that of sham operation group(P<0.01),description of ciliary neurotrophic factor on neurons after spinal cord injury is the most obviously.Chapter fourResearch Background: Pretreatment of neurotrophic factors in vitro can mobilize neural stem cells.Research aim: The aim of this study was to research findings,comparative analysis of brain-derived neurotrophic factor,ciliary neurotrophic factor,transforming growth factor score BBB,pretreatment of neural stem cell transplantation for treatment of spinal cord injury animal model to improve the effect of rotating rod balance test and slope angle test,and the pathological change of neurons degeneration the role of evaluation system of neurotrophic factor pretreatment neural stem cell transplantation for treatment of spinal cord injury treatment.Research method: Immunofluorescence detection of nerve stem cell specific marker Nestin expression verification transplantation pretreatment of cells into neural stem cells,Brd U in vivo labeling combined with immunofluorescence detection after transplantation of neural stem cell differentiation in host product neurons(beta-Tubulin III)and astrocytes(GFAP)expression,verified after transplantation of neural stem cells can survive in the host body,with biological function and differentiate into neurons and astrocytes;score test effects of different neurotrophic factors on mouse hindlimb function after spinal cord hemisection by hindlimb function BBB,screening of neurotrophic factor therapy is best;the rotary balance test and experimental test to evaluate the therapeutic effect of slope angle different neurotrophic factors on mouse behavioral function after spinal cord hemisection,selected from improving Neurotrophic factor has the strongest effect;to observe the effects of different neurotrophic factor on neurons injury of spinal cord after silver staining,were selected for follow-up study at least denatured neurons neurotrophic factor.Result and Conclusion: Immunofluorescence results showed that cell screening positive expression of Nestin and further confirmed for the study cells into neural stem cells;spinal cord slices of Brd U/ beta-Tubulin III,Brd U/GFAP double staining showed positive results that transplanted neural stem cells can survive in the host body,and differentiate into the biological functions of neurons and astrocytes BBB cells;hindlimb function scores showed that BDNF pretreatment group was significantly higher than that of ciliary neurotrophic factor(P<0.01)treatment group was higher than that of transforming growth factor(P<0.05)pretreatment group was significantly higher than that in the sham operation group(P<0.01),that brain derived business in promoting the ability of God son hindlimb functional recovery after spinal cord injury the highest;rotary balance test results show that the pretreatment of ciliary neurotrophic factor group was higher than that of BDNF pretreatment group(P<0.05)higher than that of transforminggrowth factor pretreatment group(P<0.01)was significantly higher than that of sham operation group(P<0.01).The angle of slope test test results show that BDNF pretreatment group was significantly higher than that of transforming growth factor(P<0.01)group was higher than that of ciliary neurotrophic factor(P<0.01)pretreatment group was significantly higher than that of sham operation group(P<0.01);brain derived neurotrophic factor can effectively promote the rehabilitation effect after spinal cord injury;neuron degeneration silver staining counting showed that CNTF pretreatment group was significantly lower than that of transforming growth factor pretreatment group(P<0.05)is lower than that of brain-derived neurotrophic factor(P<0.05)pretreatment group was significantly lower than that of sham operation group(P<0.01),description of ciliary neurotrophic factor on neurons after spinal cord injury is most obvious.