Effect Of Bone Marrow Mesenchymal Stem Cells Differentiate Into Neuron-like Cells On The Ability Of Learning And Memory In Cognitive Impairment Rats

Abstract Objective:Observe the effect of a multiple of the different passages of bone marrow mesenchymal stem cells (BMSCs) differentiate into neuron-like cells transplantation on the ability of learning and memory in cognitive impairment rats, and provide theoretical and experimental basis for stem cell transplantation in the degenerative diseases of the central nervous system.Methods:BMSCs harvested from healthy3-month-old male Wistar rat femur, then were passaged and purified in culture medium and cultured for5passages with adherent cultivation. The3rd passage BMSCs were tested by flow cytometry for CD29and CD44and CD34to identify the purity. Culture medium containing0.2volume fraction of fetal bovine serum and1mmol/L β-mercapto-ethanol (β-ME) were used for24-hour pre-induction. Remove the pre-induction solution,20ng/mL of bFGF induced differentiation48h. Take induced by the3rd generation BMSCs immunocytochemistry identification of neural cell-specific antigen marker neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) expression. Natural aging Wistar rats were randomly divided into four groups after the screening of the Morris water maze (n=10). Control rats with bilateral hippocampal injection of normal saline,3,4,5cells rats were injected to the neuron-like cells after induction of3,4,5-generation of BMSCs. Learning and memory in rats4weeks after transplantation and experimental determination of the Morris water maze test. Rats in each group by immunohistochemical staining comparison transplantation hippocampal choline acetyltransferase (ChAT)-positive cell number.Results:Successfully isolated and cultured Wistar rats of BMSCs, Flow cytometer studies presented that the cultured BMSCs were CD29,CD44positive and CD34negative, in line with BMSCs phenotypic characteristics. The majority of3,4,5generation BMSCs after induction can differentiated into neuron-like cells, the3rd generation BMSCs induced immunocytochemistry identification of neural cell-specific antigen markers NSE positive expression, negative expression of GFAP. Morris water maze test results:compared with pre-transplant, learning and memory decline in control rats, but the difference was not significant (P>0.05);3,4,5cells rats learning and memory are increased (P<0.05). Compared with the control group,3,4,5-generation cell group in rats learning and memory were improved (P<0.05). 3,4,5-generation cell group pairwise comparisons, the differences were significant (P<0.05).After transplantation, the rats hippocampal ChAT positive results: compared with the controlgroup,3,4,5-generation cell group in rat hippocampus ChAT positive cell counts wereincreased (P<0.05).3,4,5-generation cell group pairwise comparison, the difference was notsignificant (P>0.05).Conclusion:3,4,5-generation rat BMSCs differentiate into the neuron-like cells transplantationcan improve learning and memory ability of cognitive impairment rats, and the4th generationcell transplantation is superior to3and5generations.