Optimization Methods About Primary Culture of Hippocampal Neurons and Purity Identification

Objective:To explore a simple and high-purity method about primary culture of rat embryonic hippocampal neurons,through optimization and improvement of traditional neuron culture.Methods:Pregnant rats were sacrificed by cervical dislocation,and the rat embryonic hippocampal tissue was taken out quickly.The hippocampal neurons were separated,digested and made into cell suspension,and then inoculated into DMEM/HG medium at an appropriate density.DMEM/ HG medium were replaced by neurobasal medium after 4hours.Thereafter,one third volume of culture medium was refreshed every two or three days.The neurons' growth and morphological changes were observed with inverted microscope,neurons activity was detected with CCK-8Kit,and the purity was identified with neuron-specific enolase(NSE).Results:Hippocampal neurons had adhered after being cultured for 4hours.Neurons had a series of morphologic changes with extending of incubation time.The purity of neurons increased gradually with extending culture time,and it was over 90percent on the 7th day.Neurons were matured on the 9th or 10th day and were used for the follow-up studies.Conclusion:The serum-free culture method is simple to acquire high purity of neurons,cell activity is good,and cell survival time is long in vitro.It is worth being applied widely in vitro studies as a good cell model.