An optimized culture method of primary hippocampal neurons in SD rats / 中国药理学通报

ABSTRACT

Aim To establish a stable and efficient method for the primary culture of hippocampal neurons from serum-free fetal rats.so as to obtain hippoeampal neurons with higher purity and better vitality.Methods The hippocampal tissues of SI) rat fe¬tus rats on the ( 18 ± 1) day of pregnancy were cultured by Neu- robasal or DMEM/F12 medium and divided into serum-free cul-ture group, fetal bovine serum culture group and 5-fluoro-2'de- oxyuridine culture group.After cells of three groups were cul¬tured for 12 hours, all medium was changed to Neurobasal medi¬um.When the neurons in FUI)R culture group were cultured to 3 days.FLDR was added to inhibit the growth of glial cells.'Hie growing status of hippo cam pal neurons at different culture time points was observed,neuronal activity was detected,cell apopto¬sis was assessed, and the purity of hippocampal neurons was i- dentified.Results Compared with 10% serum culture group, the neurons cultured in serum-free culture group showed higher viability,lower apoptosis rate and higher purity; FUI)R reduced cell viability and increased cell apoptosis.Conclusions 'Hie neurons cultured by serum-free culture have good activity and high purity, instead of adding glial cell inhibitors, which is a fea¬sible and optimized primary culture method for hippocampal neu¬rons.