Optimization of transduction conditions of recombinant adeno-associated virus into NK92 cells
10.11665/j.issn.1000-5048.20221109001
- VernacularTitle:重组腺相关病毒对NK92细胞的转导条件优化
- Author:
Xiaoli SHEN
1
;
Qungang ZHOU
;
Ran ZHENG
;
Xiangjun SHEN
;
Xiang QIU
;
Taiming LI
Author Information
1. 中国药科大学生命科学与技术学院
- Publication Type:Journal Article
- Keywords:
NK92 cells;
adeno-associated virus;
transduction efficiency
- From:
Journal of China Pharmaceutical University
2023;54(1):115-121
- CountryChina
- Language:Chinese
-
Abstract:
To improve the transduction efficiency of recombinant adeno-associated virus (rAAV) in NK92 cells, the number of cells, concentration of IL-2 in the medium, and serotype and dosage of rAAV were explored to optimize cell state and viral transfection conditions.Then, zinc chloride (ZnCl2), chloroquine, polyvinyl alcohol (PVA) and genistein with different concentration were added separately during transfection to further improve the viral transduction efficiency.The results showed that, at cell number of 5 × 105, the expression efficiency of enhanced green fluorescent protein (EGFP) was relatively high.When the IL-2 concentration was 1 000 IU/mL, NK92 cells were most suitable for virus transfection. The transduction efficiency of different serotypes of rAAV in NK92 cells was rAAV6, rAAV2 and rAAV9 in descending order.Pretreatment of NK92 cells with genistein could significantly increase the viral transduction efficiency, while the addition of other reagents had no significant effect.Through the optimization of the above conditions, the transduction efficiency of rAAV to NK92 cells could be significantly improved, which provided evidence for functional genetic modification of NK92 cells by rAAV.
