A Cultural Method of Hippocampal Neurons of Rats

Objective To establish a efficient method of primary hippocampal neurons.MethodsThe hippocampus were harvested from SD rat(during 24 h) under sterile conditions,rinsed and cut to shiver,digested by 0.25%trypsin,filtered with 200 Sieve,mixed culture.Cytosine arabinoside(Ara-c) was used to inhibit the development of glial cells after cultivated for 5 days.The purity of isolated cells were identified by NSE immunocytochemical staining.Results More than 90% of Primary hippocampal neurons was obtained by this method.A typical neuronal morphology appeared after cultivated for 3 days,up to 13th or 14th day,many neuritis extended to form intensive network,at last,cell fragment was obsevered in 14 days later.Conclusion The cultural method is a convenient method with higher activity and purity,providing a basis for researching the related disorders of the hippocampal neurons.