Abstract: Objective: To establish an ideal method for primary culture of neurons from cerebral cortex of neonate rats.Methods: The cerebral cortex of newborn rats was digested with trypsin to obtain single cell suspension,after the cell viability was estimated by trypan blue staining,the cells were planted;when the cells were cultured for 3-4 d,Ara-c was added to purify the neurons;the cultured cells were finally,identified by toluidine blue stain for Nissl bodies.Results: The survival rate of the conducted cells was high;under the culture condition in vitro,the cells grew well.When the cells grew for 6-7 d,the cell body was big,the prominences were thick and there were many branches;the prominences interlaced to net,forming typical nerve fiber net.When the cells grew for 10-14 d,cells with Nissl bodies were above 90%.Conclusions: It is an ideal technique for culture of cortical neurons from cerebral cortex of neonate rats in vitro.