Functional Development Of Potassium Current In Neural Stem Cells Cultured In Vitro

It is shown recently that neural stem cells(NSCs) are widely distributed in embryonic and adult brain, and peripheral nervous system. Furthermore , the NSCs can be induced into neurons,astroglias and oligodendrocytes by cytokines to compensate for the damaged nervous tissues. However, it is not clear whether NSCs cultured in vitro are fit for transplantion therapies. To discuss the feasibility of the cells transplantion, it is important to ensure that NSCs must be functional after differentiation in vitro. However, the current research is limited to immunocytochemistric method to identify molecular markers of differentiated NSCs, while the excitable features get no enough attention. Therefore, in this study, voltage patch clamp technique was applied to detect the ion current in different differentiation terms, which, combinded with the data of protein markers, will help to identify the type and function of induced NSCs, and to provide theoretic and experimental foundation for clinical application of NSCs.MethodsNSCs were isolate and culture from dentate gyrus of hippocampus in newborn rat . After 3-4 passages in vitro, single cells from neurospheres was induced to differentiate by adding 10% Fetal bovine serum and simultaneous removal of mitogens. Before and after differentiation, protein markers in the cells were identified separately with anti-nestin , anti-NSE and anti-GFAP antibodies...