26 October 2011
Neurotrophin-3 gene transduction of mouse neural stem cells promotes proliferation and neuronal differentiation in organotypic hippocampal slice cultures
Hai-xia LuACDEFG, Zhi-ming HaoACDEF, Qian JiaoBCF, Wu-ling XieBC, Jun-feng ZhangBF, Yi-fei LuB, Min CaiB, Terry ParkerDE, Yong LiuADGDOI: 10.12659/MSM.882039
Med Sci Monit 2011; 17(11): BR305-311
Abstract
Background: The transplantation of neural stem cells (NSCs) has been accepted as a promising therapeutic strategy for central nervous system disorders. However, the beneficial effect of NSC transplantation upon functional recovery is limited due to the unfavorable microenvironment (niche) at the site of trauma or degenerative disease in the brain. Combination of transplantation of NSCs with neurotrophins may overcome the hurdles of impaired cell survival and neuronal differentiation.
Material/Methods: In the current study, the neurotrophin-3 (NT-3) gene was transduced into cultured mouse embryonic cortical NSCs via an AAV vector (NSC-NT-3). The effect of NT-3 over-expression on cell proliferation and differentiation in NSCs was observed by immunohistochemistry, cell culture and organotypic hippocampal slice cultures.
Results: The characteristics of self-renewal and multiple differentiation of NSCs were well-preserved. Cells in the NSC-NT-3 group proliferated faster and differentiated into more -tubulin III-positive neurons compared to the control group in vitro. Furthermore, cells in the NSC-NT-3 group survived in a significantly higher percentage and undertook neuronal differentiation preferably in organotypic hippocampal slice cultures.
Conclusions: Our results suggest that the transduction of NT-3 into NSCs could effectively promote NSCs survival, proliferation, and neuronal differentiation in vitro without change of the stemness of NSCs. This work also offers evidence to better understand the safety and efficiency of combined treatment with NT-3 and NSCs for the central nervous system disorders.
Keywords: Neural Stem Cells - transplantation, Hippocampus - cytology, Genetic Vectors - genetics, Enzyme-Linked Immunosorbent Assay, Dependovirus, Cell Differentiation - physiology, Analysis of Variance, Neurotrophin 3 - metabolism, Transduction, Genetic - methods
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