Optimization of culture method of mouse primary hippocampal neurons and construction of HT22-GRK2 －/ － cells
10.19405/j.cnki.issn1000-1492.2023.04.012
- Author:
Menghui Guo
1
;
Nana Xue
1
;
Xi Yuan
1
;
Qian Meng
1
;
Wei Wei
1
Author Information
1. Institute of Clinical Pharmacology，Anhui Medical University，Key Laboratory of Anti-inflammatory and Immune Medicine，Ministry of Education，Anhui Collaborative Innovation Center of Anti-inflammatory and Immuno Drugs，Hefei 230032
- Publication Type:Journal Article
- Keywords:
hippocampal neurons;
primary culture;
HT22;
GRK2;
CRISPR / Cas9
- From:
Acta Universitatis Medicinalis Anhui
2023;58(4):589-596
- CountryChina
- Language:Chinese
-
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.
- Full text:2024071710533446099小鼠原代海马神经元培养方法...RK2_(-_-)细胞构建_郭梦慧.pdf
