Optimization of culture method of mouse primary hippocampal neurons and construction of HT22-GRK2 -/ - cells / 安徽医科大学学报
Acta Universitatis Medicinalis Anhui
; (6): 589-596, 2023.
Article
en Zh
| WPRIM
| ID: wpr-1038506
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WPRO
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.
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Acta Universitatis Medicinalis Anhui
Año:
2023
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Article