Abstract
Objective To investigate the effect of blocking hairy and enhancer of split 1 (Hes1) on gamma aminobutyric acid (GABA) neurons’ differentiation in human neural stem cells. Methods The human neural stem cells were cultured for 10 days and identified by nestin immunofluorescence stain, then using the human neural stem cells to differentiate GABA neurons. The design project of experiment was adopted completely random. The control cultured medium were the common differentiated medium added 10% bovine serum albumin. And the experimental cultured mediums were identical with the control excepted for the anti-Hes1. Anti-GABA immunofluorescence stain was carried out when most of the cells were differentiated. Results A lot of 150-200 μm diameter neurosphere which were identified when cultured for 10 days. The second generation of neurospheres were using for GABA neurons differentiation. At the sixth day, more than half of cells were differentiated and begin to concentrate. Digested and inoculated again, then 90%-95% cells were adherent and differentiated at day 14. Anti-GABA immuno-fluorescence stain showed that 55-70 positive cells were be found in 100 cells in every visual field, the rate was (64.0±0.6)% in anti-Hes1 differentiated group and in the same condition the positive cells were only 15-20 positive cells, the rat was (16.6±0.6)% in control group, there are significant difference between the two groups. Conclusion blocking Hes1 can significant increase the differentiation rate of GABA neurons in human neural stem cells. Key words: Hes1; Human neural stem cell; Gamma aminobutyric acid ergic neuron; Differentiation
Published Version
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