Objective To isolate, cultivate and identify the neural stem cells (NSCs)of mouse embryo in vitro and study the effect of paraqua(t PQ)on their proliferation viability.
Methods Cerebral cortexes were isolated from embryonic (E10.5 d) mice. The cells were identified as neural stem cells through immunocytochemistry assey with antibodies to neural stem cells (NSCs) specific protein and morphological observation. After treatment with 0.00, 0.10, 1.00, 10.00, 100.00, 1 000.00 μmol/L PQ for 24 h, the viability of the NSCs was measured by Alamar Blue assay. After treatment with 0.00, 0.10, 1.00, 10.00, 50.00, 100.00μmol/L PQ for 24 h, the reactive oxidative spieces (ROS)production of the NSCs was measured by DCFH-DA assay.
Results The result of immunocytochemistry assey showed that nestin which is the specific protein of NSCs was expressed in the majority of the cells. Alamar Blue assey reveals that the cell viability was reduced significantly in the groups of PQ ≥ 100.00 μmol/L compared with the control group (P < 0.05). With the increase concentration of PQ, the cell viability decreased accordingly (P < 0.05). DCFH-DA result showed that the ROS production increased significantly in the groups received PQ ≥ 10.00 μmol/L compared with the control group (P < 0.05). Concentration with the increase of PQ, the ROS production increased accordingly (P < 0.05).
Conclusion Primary culture of NSCs in vitro was established successfully. PQ contributed a dose-dependent relationship on suppression of the proliferation and raise of ROS level of NSCs.
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