Purification and identification of hippocampal neurons primary cultured in vitro

Objective To establish an optimizing method of purification and identification of hippocampal neurons primary cultured in vitro.Methods Hippocampus was disconnected from new-born rat and then hippocamal neurons were cultured with both serum medium and serum-free medium in vitro.Identification and purification of neurons was detected with specific NSE、GAP-43 and MAP2 antibodies by immunocytochemistry. Results All cells adhered to the footwall after 24 hours and short nervous processes were found in subtotal neurons,and these processes extended markedly with time and connected into nets.Cells were fully developed on the 7th～8th day,and then began aging;4 weeks were the longest life span observed.Hippocampal neurons got more than 96%purity after identifying.Conclusion Primary culture of hippocampal neurons with serum medium and serum-free medium in vitro is with high purity and low heterogeneity,as can be used as cell basis in research of nervous diseases in vitro.