Isolation, cultivation and identification of hippocampal neurons from prenatal mice

Objective To establish a favorable primary culture of hippocampal neurons isolated from prenatal mice. Methods The hippocampus of embryonic SD mouse aged 18 d was isolated, digested with papain, and then the neurons were cultured in planks with polylysine before experiment, we used Neurobasal as a culture medium for neurons which was without serum. And the specific markers of neurons were identified through immunofluorescence using MTT colorimetric analysis to detect the survival rate of neuronal cells under different treatments. Results Hippocampal neurons grow well, synapses grow out after 24 hours from cultivation and a mature network of nerve cells can be formed after 7-12 d. The NSE immunofluorescence identified neurons of more than 95% purity. Compared with trypsin, the cell survival rate was higher with papain in digestion (P<0.01); The cell survival rate of normal tip -1.0 mm was higher than tip-0.5 mm and 200 mesh sieve (P<0.01). Conclusions By this technique, we can obtain hippocampal neurons with high purity, and it is a simple, effective method for cell culture. Key words: Hippocampal neurons; Primary culture; Prenatal SD mice