The study of hypoxia tolerance of hippocampal neurons from 5-day-old rats

Objective To establish a primary culture technique for 5-day-old rat hippocampal neurons and investigate the hypoxia tolerance of this kind of neurons.Methods To improve primary culture technique suitable for 5 days old rat hippocampal neurons,the neurons were divided into the oxygenation group and control group.Inverted microscope and immunohistochemistry of NF were performed to assess the growth and morphology of the cells.The neurons from 5-day-old and 1-day-old rats were exposed to hypoxia condition(0%02,5%CO_2 and 95%N_2)for 12h or 24h after 1 week primary culture,respectively.MTT cell assays measured cell viability.Results(1)The growth ofhippocampal neurons from 5-day-old rats in the oxygenation group was more vigorous than in the control group.(2) The cell viability of neurons from 1-day-old rat and 5-day-old rats with 12 h hypoxia exposure were (81.5±2.3)%(n=8)and(89.4±2.2)%(n=6)(P 0.01)respectively,while that with 24 h hypoxia exposure were(75.0±2.8)%(n=8)and(85.0±2.2)%(n=6)(P0.01)respectively.During hypoxia,the hippocampal neurons disassociated from 5-day-old rats were more tolerant than that from 1-day-old rats.Conclusion The oxygenation during dissection and culture of hippocampal neurons was beneficial to the growth of neurons,and the hypoxia tolerance of 5-day-old rats hippocampal neurons were better than the that of 1-day-old neurons.