Experimental Study Of Neural Stem Cell Transplantation In The Brain Of Alzheimer’s Disease

Objective:With the development of living standards and the changing of our social environment, the incidence rate of Alzheimer disease (AD) presents a rapidly rising trend。According to medical studies, Alzheimer’s disease has not yet have the effects of drugs and treatment methods. In this experiment, the green fluorescent protein (GFP) transgenic mouse embryonic neural stem cells (NSCs) were isolated, cultured and purified.After then transplanted into the hippocampus of APP/PS1double transgenic mouse brain. TO explore the neural stem cells repaired in the process of rat brain. Providing a theoretical basis for the clinical application of neural stem cells which treat Alzheimer’s disease.Methods:(1)The GFP transgenic mouse embryos of E14, Isolated from their brain tissue which made of nerve stem cells in suspension culture. Immunofluorescence was used to detect the Nestin.Fetal bovine serum induced neural stem cell differentiation.The MAP-2(neuron-specific enolase enzyme)and GFAP (glial fibrillary acidic protein) immunocytochemical staining were Identificated of neurons and astrocytes.(2) E14of the C57mice embryos using the above method of neural stem cell suspension culture. The passage4-6generation density of each well8×104-1×105cells/of500μl were seeded in24-well plates for0,1,5,10,15,20exposed, with the multiplicities of infection(MOI), lentiviral vector coding for the GFP gene (LV/GFP) diluent, six wells seeded for the degree of a drop, a total of six groups.2-3days later placed under inverted fluorescence microscope to observe the positive expression of GFP.Examination by flow cytometry in each group of neural stem cells,GFP-positive transfection rate.(3) The neural stem cell of GFP transgenic mouse transplantation in AD mouse hippocampus. After A week, do a water maze and open field test to detect changes in learning and memory ability of the mice. After four weeks, the detection of neural stem cell survived and migrated. Results:The in vitro experiments show that the neural stem cells cultured from the separation of E14fetal rat brain, And the vast majority of the cells can express nestin. Induced by fetal bovine serum differentiation medium, prove the potential of multi-directional differentiation of neural stem cells. A small number of the stem cells can differentiate into MAP-2expression in neurons, the majority can be differentiated into the expression of GFAP by astrocytescells.Neural stem cells transduction with the GFP gene, After2-3days,each well GFP expressing except in the control group. With the increase of the MOI, the positive expression rate increased. MOI=0-10, P<0.05.Learning and memory testing, Cell transplantation group have great differences than model group (p<0.05).There was no significant difference compared with control group.Immunohistochemistry to detect:After transplantation of neural stem cells can not only integrate with AD micebrain tissue, but also migrate within the AD brain tissue of mice Conclusion:Neural stem cells in serum-free complete medium culture survival and proliferation. Migration and integration of the transplanted hippocampal tissue can be completed. Neural stem cells in the treatment of Alzheimer’s disease have a certain effect. Lentivirus can be used as the exogenous gene into neural stem cells idealvectors.