The Therapeutic Effect Of Various Neural Stem Cells On Brain Contusion And Laceration In Cloning Goats

The regeneration of nervous tissue in central nervous system is a tough problemin medical science. Neural Stem Cell(NSC)transplantation serves as a newly emergingand a good perspective therapeutic method, has been confirmed in many diseasetherapy in neuroscience domain. Brain trauma is the most frequent disease in clinicalneurosurgery and has largest harm to mankind. It has the topmost therapeutic value.Even though different brain trauma has different pathological process, but most ofthem have the brain contusion and laceration. So We researched the brain contusionand laceration restrictly in our study.Based on its resource, NSC can be divided into two groups: embryonic derivedneural stem cell(EdNSC)and adult derived neural stem cell(AdNSC). Whosetherapeutic effect on brain contusion and laceration is better? Do immunologicalrejection generate When various kinds of NSC transplanted into brain of animal? Canestablished a brain contusion and laceration model in big animals that have highsurvival rate and can be detected clearly? To these problems, there has no convinceresearch now. In our study, We utilize the cloning goats Whose heredity backgroundare very clearly as the models to estabilsh the brain contusion and laceration ofcloning goats, and investigate the immunological rejection of EdNSC and AdNSCtransplantation.In the first part of our study, We obtained the cerebral cortex of adult cloninggoat(NO:CL-4) and non-cloning goat(NO:SP-3119) by surgical intervention. Weobtained the AdNSC of them by cell culture in vitro. Then We obtained the cerebralcortex of the fetal cloning goat(35days)Which gained by somatic cell nucleartransfer(SCNT) of CL-4 cloning goat and obtained the EdNSC by cell culture in vitro.To the CL series cloning goats,the EdNSC and AdNSC of cloning goat could be deemas "autoallergic NSC", and AdNSC of non-cloning goat was alloallergic NSC.On theother hand, in the cell culture process, the EdNSC growed faster than AdNSC in thesame cell culture method.In the second part of our study, We chosed 20 CL series cloning goats at random.After exposed the upper Sylvii sulcus moter cortex of brain of the cloning goats bysurgical intervention, 50 gram of steel ball was used to produce a free fallingaccelerate strike on the brain from 100cm height respectively and established thebrain contusion and laceration goat models that had high survival rate and could be detected clearly.In the third part of our study, We divided the cloning goat models into 4 groups.We transplanted the EdNSC, AdNSC of cloning goat and AdNSC of non-cloning goatand saline into the around areas of brain contusion and laceration of each groupmodels respectively. We observed each group models' survival rate, ethologicalrecovery; The Motion Evoked Potential(MEP), brain Magnetic ResonanceImaging(MRI) image, pathology changes of brain contusion and laceration in eachgroup models were also recorded. We then observed the survival, immigration anddifferentiation of all kinds of stem cells in goats' brains. We found: all kinds of stemcells can survival, immigrate and different well in the transplanted brain. The EdNSCand AdNSC of cloning goat transplantation groups had good survival rate, ethologyrecovery. Their MEP recovery of the motor cortex, brain MRI image and pathologychanges of brain contusion and laceration were better than AdNSC of non-cloninggoat transplantation group and control group; The EdNSC transplantation group hadthe best therapeutic effect on brain contusion and laceration in all transplantationgroups and control group.In the fourth part of our study, We investigated the acute(1 week after celltransplantation) and chronic(3 months after cell transplantation) immunologicalrejection of each group of the models Who received various transplanted stem cells.We detected the various cytokine(IL-2,IL-6 and IL-10) in each group models' blood atdifferent time (1 day before model established; before, 3 days, 1 month and 3 monthsafter cell transplantation).We also detected CD3+ cells in the cell transplantation areasaround brain contusion and laceration in all groups goats by the method ofimmunohistochemistry. We found:in the AdNSC of non-cloning transplantationgroup,the level of IL-2 was obviously high and the level of IL- 10 was obviously lowthan other groups at 3 days, 1 month, and 3 months after celltransplantation(P＜0.05).The level of IL-6 was obviously high than other groups at 3months after cell transplantation(P＜0.05). The level of these cytokines in the EdNSC,AdNSC of cloning goat transplantation groups and control group had no statisticsdifference(P＞0.05) at every time. The quantity of CD3+ cells in the AdNSC ofnon-cloning goat group was much more than that of other groups(P＜0.05) at the acuteperiod(1 week after cell transplantation) and chronic period(3 months after celltransplantation). The quantity of CD3+ cells in the EdNSC, AdNSC of cloning goattransplantation group and control group had no statistics difference(P＞0.05) at acute and chronic period.Overall, in our study, We got the conclusions as follow: 1,The EdNSC, AdNSCof cloning goat and the AdNSC of the non-cloning goat were successful obtained;EdNSC growed faster than AdNSC in the same cell culture method. 2,The braincontusion and laceration model of cloning goats were successful established by freefalling of 50 gram of steel ball accelerate strike on the goat brain from 100cm height.3, EdNSC and AdNSC transplantation both had obvious therapeutic effect on braincontusion and laceration of cloning goats; The therapeutic effect of EdNSCtransplantation was better. 4, Allograft of stem cells has acute and chronicimmunological rejection and autografting of stem cells has no acute and chronicimmunological rejection.