Effects of non-essential amino acids andβ-mercaptoethanol on proliferation of neural stem cells

WANG Weiwei; BAO Tianhao; ZHANG Hongmiao; WANG Shufen; XIAO Zhicheng; HAN Jianhong

doi:10.3969/j.issn.1002-266X.2016.01.007

Abstract：Objective To investigate the effects of combined application of non-essential amino acids ( NEAA) and β-mercaptoethanol on proliferation of neural stem cells (NSCs).Methods The female rats in the gestational age of 14.5 d were selected and executed to extract NSCs and cultivate.We took 5-7 generations of NSCs to do the experiments.The NSCs cultured were divided into four groups:the control group which was added with 12μL phosphate buffer solution (PBS), the NEAA group which was added with 10 L NEAA and 2 L PBS, theβ-mercaptoethanol group which was added with 2μLβ-mercaptoethanol and 10 L PBS, and the NEAA +β-mercaptoethanol group which was added with 10 L NEAA and 2μLβ-mercaptoethanol.The pro-liferation of NSCs was calculated by cytometry, the proliferation rate of NSCs was detected by MTT, the caspase-3 expression of NSCs was assayed by RT-PCR, and NSCs were identified and examined for the differentiation potential of NSCs into neurons and astrocytes by immunohistochemistry.Results Compared with the control group, the proliferation speed and rate of NSCs in the NEAA group were increased, but the caspase-3 mRNA expression of NSCs was decreased (P<0.05 or P<0.01);the prolifera-tion speed and rate of NSCs were increased, but the caspase-3 mRNA expression of NSCs, the percentage of positive ATUJ-1 cells, the percentage of positive GFAP cells were decreased and Nestin was increased in theβ-mercaptoethanol group and NEAA+β-mercaptoethanol group (P<0.05 or P<0.01).Significant difference was found in each index between the NEAA group and NEAA+β-mercaptoethanol group (P<0.05 or P<0.01).Compared with the β-mercaptoethanol group, the proliferation speed and rate of NSCs were increased, but the caspase-3 mRNA expression of NSCs was decreased in the NEAA+β-mercaptoethanol group (P<0.05 or P<0.01)Conclusion Both NEAA andβ-mercaptoethanol can promote the proliferation of neural stem cells and they have synergistic effect during the combined application.