Optimization of transduction conditions of recombinant adeno-associated virus into NK92 cells

SHEN Xiaoli; ZHOU Qungang; ZHENG Ran; SHEN Xiangjun; QIU Xiang; LI Taiming

doi:10.11665/j.issn.1000-5048.20221109001

Journal of China Pharmaceutical University > 2023 > 54(1): 115-121. > DOI: 10.11665/j.issn.1000-5048.20221109001

SHEN Xiaoli, ZHOU Qungang, ZHENG Ran, SHEN Xiangjun, QIU Xiang, LI Taiming. Optimization of transduction conditions of recombinant adeno-associated virus into NK92 cells[J]. Journal of China Pharmaceutical University, 2023, 54(1): 115-121. DOI: 10.11665/j.issn.1000-5048.20221109001

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Optimization of transduction conditions of recombinant adeno-associated virus into NK92 cells

1.
School of Life Science and Technology, China Pharmaceutical University, Nanjing 211198
2.
Suzhou Blood Center, Suzhou 215005, China

Funds: This study was supported by Suzhou Municipal Science and Technology Program (No.GWZX201903)

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Received Date: November 08, 2022
Revised Date: February 21, 2023

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Abstract

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.
- NK92 cells,
- adeno-associated virus,
- transduction efficiency

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