The Experimental Research Of Neural Stem Cells Isolation Culture And Transplant To Treat Rat Cerebral Hemorrhage

The cells were isolated from adult Individual,it have the ability of multi-direction differentiation and self-refresh,named the neural stem cell.Hence surmounted the old concept of adult mammalia animals center nervers system usually were thinked constitution of cannot regeneration. Objective:1,To establish the perfection methods of isolation,culturing and evaluation of neural stem cell.,2,Trying use the Neural stem cells transplant to treat cerebral hemorrhage. Methods:The advantage of serum free culturing technology was performed to islate from 14-16days embrynic rat hippocampas,striatum and culturing with bFGF(basic fibroblast growth factor).Differentiation cultur with 5% embrynic bovine blood serum and exclude growth factor in vitro. Fluorescence Immunocytochemica were applied to identify the neural stem cells and the cells of differentiated. The rats intracerebral hemorrhagerat models were made。Animals were randomly divided to four groups control group,three days transplant group after intracerebral hemorrhagerat,seven days transplant group after intracerebral hemorrhagerat and bFGF injection group.the neural stem cells or bFGF were transplanted into the caudate nucleus 7days later.The motor function of rats were investigated at 2hours and four week after transllantion.Than the rats were sacrificed.The condition of growth and ｍigration of neural stem cells invivo were checked. Results:the cell clones isolated from embrynic rat hippocampus,striatum possess the ability of proliferation,self-reproduction.Nestin positive cells can be found. Through microscop the cell clones showed typical characteristics of neural stem cell. Immunocytochemica were applied to identify the Produce of differentiation:neur cell,glial cell and oligodendrocyte.The IgG antibody Microtubule-associated protein can identification neur,the IgG antibody of Glial fibrillary acidic protein can identification glial cell,the IgG antibody of Cyclic nucleotide phosphodiesterase can identification oligodendrocyte.The result of experiment display:Four groups animal gain recovery.The motor function of rats that recieved transplanting 7days after cerebral hemorrhage recovered much better than the 3days group,bFGF group and the control group. Brain constitution pathological section can find surviver cells that had were transplantationed and the neural stem cells self-body. Conclusions:The neural stem cells are a sort of primitive precursor cell be un-differentiation bihavior.It posses three behavior self-refresh,low immune response,multi-direction differentiation potential.Neural stem cells are very appropriated for transplantation after damage of nervous tissue,to investigation neural stem cells transplantation repair and replacement nervous tissue after cerebral hemorrhage had damaged and function recovery have very important themry significance and utility value.Various regions of the adult body brain, including the hippocampus, the ventricular/ependymal zone, the corpus striatum existence neural stem cells,if can induce them proliferation differentiation to replace the deficient nervous tissue with disease that would provid new hope for injury nerver. Various locus and sorts neuronal regeneration differentiation have its special character,demand various cell facter to control strictly,thus it is a important content of neural stem cells to clarification various cell factor's regulation in brain.Though can not accurate induce neural stem cells specific orientation differentiation yet,to accompany the unceasing deepen recognize of convection mechanisms in cells' signal convection and discovery of new cell facter,new chemo-signal, Homo sapiens would solve the question of orientation differentiation from totipotential stem cell,multipotential stem cell to neural stem cell, orientation nerver ancestor cell then special tipe neural cell, implementation functional reconstruction betwen neural cell,for ultimate solve the proble of neural regeneration developing the way.