Optimization of culture method of mouse primary hippocampal neurons and construction of HT22-GRK2 －/ － cells / 安徽医科大学学报

ABSTRACT

Objective@#To explore and optimize the primary culture method of neonatal mouse hippocampal neurons in vitro．To construct a G-protein-coupled receptor kinase 2 ( GRK2) knockout HT22 cell line．@*Methods @#Hippocampal tissue of C57BL6 /J mice on day 1－2 was taken，digested with trypsin and pipetted to form a cell suspension，and supplement was added to Neurobasal-A medium to maintain cell growth． CRSIPR / Cas9 gene editing technique was used to construct HT22-GRK2 －/ － cell line，and the knockout efficiency of GRK2 was detected by immunofluorescence staining and Western blot. @*Results @#Primary hippocampal neurons of newborn mice were put into six-well plates with 3 × 107 /well using a serum-free culture method，which could get a high purity and good activity ; HT22-GRK2 －/ － cell line was constructed successfully．@*Conclusion@#The primary culture method of mouse hippocampal neurons was successfully established and optimized，and HT22-GRK2 －/ － cell line was successfully constructed by CRSIPR / Cas9 gene editing technique.