Neural Stem Cells Combined With A Variety Of Cytokines And Fibrin Scaffold Materials Improve The Cognitive And Memory Impairment Of Alzheimer's Disease Mice

Alzheimer's disease(AD)is a neurodegenerative disease that seriously affects the health of the elderly.Its onset is mainly due to multiple neuron apoptosis caused by a large number of A? deposition and Tau protein phosphorylation,eventually lead to serious impairments of cognitive and memory.To investigate the improvement of memory function and the differentiation ability after transplantation NSCs combined with multiple factor and fibrin to hippocampus in AD model mice.Take gestational age of 10 days C57BL/6J pregnant rats,and isolate the fetal rat NSCs;In vitro differentiation and identification of NSCs;AD model mice were divided into three groups: PBS(control)group,multiple factor group and NSCs + multiple factor group.Morris water maze(MWM)and novel object recognition task were used to detect the changes of memory behavior before and after transplanting NPC cells in AD model mice;Immunofluorescence,immunohistochemistry and Western-blotting were used to detect the differentiation and migration ability of NSCs transplanted into hippocampus.Graph Pad Prism7.0 software was used for statistical analysis and groups were compared using group F test.Results show that,the isolated hippocampal neural stem cells can normally proliferate in vitro.After 14 days of in vitro differentiation,the results of immunofluorescence staining showed that the NSCs differentiated into neurons with the cholinergic neuron marker Ch AT.the memory cognitive function of transplanted cells and factors AD mice was improved at an early stage.Over time,the memory improvement of the transplanted NSCs group lasted longer.In the Morris water maze experiment,the escape latency 14.12 ± 7.45 s before the platform of the group injected with NSCs and factors was found to be significantly lower than that of the AD model mice injected with PBS 39.65 ± 4.64 s,(F = 2.578,P = 0.0094).After 24 hours with platform removed,the escape latency of the group injected with NSCs and factors 15.12±3.52 s was still lower than that of PBS group 37.17±2.18 s,(F=2.598,P=0.0003),and the time of factors group 16.62±3.23 s was also lower than that of PBS group 37.17±2.18 s,(F=2.186,P=0.0004).In the new object recognition experiment,the NSCs + factors group data 68.46 ± 2.4% was higher than PBS group 54.47 ± 4.79%,(F = 3.983,P = 0.018).The factors group 65.20 ± 1.03% was also higher than PBS group 54.47 ± 4.79%,(F = 21.63,P = 0.042).Western-blotting results showed a decrease in cholinergic neurons in the hippocampus of AD model mice compared with normal C57 mice.After transplantation of NSCs,the number of cholinergic neurons in the brain of AD model increased.Immunofluorescence and immunohistochemistry showed that,the transplanted NSCs survived in the brain and differentiated into cholinergic neurons.NSCs combined with multiple factor and fibrin transplanted in bilateral hippocampus of AD model mice improve memory cognitive function of AD mice by differentiating into functional cholinergic neurons.