Enhanced non-viral gene delivery via calcium phosphate/DNA co-precipitates with low-voltage pulse electroporation in NK-92 cells for immunocellular therapy

Che-Yung Kuan; I-Hsuan Yang; Chia-Ting Chang; Zhi-Yu Chen; Jhih-Ni Lin; Wei-Ting Kuo; Yu-Ying Lin; Andrew Yueh; Feng-Huei Lin

doi:10.1063/5.0198191

Enhanced non-viral gene delivery via calcium phosphate/DNA co-precipitates with low-voltage pulse electroporation in NK-92 cells for immunocellular therapy

APL Bioeng. 2024 Aug 6;8(3):036107. doi: 10.1063/5.0198191. eCollection 2024 Sep.

Authors

Che-Yung Kuan, I-Hsuan Yang¹, Chia-Ting Chang, Zhi-Yu Chen¹, Jhih-Ni Lin², Wei-Ting Kuo², Yu-Ying Lin, Andrew Yueh³, Feng-Huei Lin

Affiliations

¹ Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, No. 35, Keyan Road, Zhunan, Miaoli 35053, Taiwan.
² Department of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, No. 49, Fanglan Rd., Taipei 10672, Taiwan.
³ Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, No. 35, Keyan Road, Zhunan, Miaoli 35053, Taiwan.

Abstract

Achieving high cell transfection efficiency is essential for various cell types in numerous disease applications. However, the efficient introduction of genes into natural killer (NK) cells remains a challenge. In this study, we proposed a design strategy for delivering exogenous genes into the NK cell line, NK-92, using a modified non-viral gene transfection method. Calcium phosphate/DNA nanoparticles (pDNA-CaP NPs) were prepared using co-precipitation methods and combined with low-voltage pulse electroporation to facilitate NK-92 transfection. The results demonstrated that the developed pDNA-CaP NPs exhibited a uniform diameter of approximately 393.9 nm, a DNA entrapment efficiency of 65.8%, and a loading capacity of 15.9%. Furthermore, at three days post-transfection, both the transfection efficiency and cell viability of NK-92 were significantly improved compared to standalone plasmid DNA (pDNA) electroporation or solely relying on the endocytosis pathway of pDNA-CaP NPs. This study provides valuable insights into a novel approach that combines calcium phosphate nanoparticles with low-voltage electroporation for gene delivery into NK-92 cells, offering potential advancements in cell therapy.