Application of orthogonal design in optimization of the transfection efficiency of polyethylenimine mediated gene transfer to hepatoma carcinoma cells.
- Author:
Yanfang ZHOU
1
;
Xiaoai CHEN
;
Meihong YE
;
Xintao SHUAI
;
Yubin DENG
Author Information
1. Department of Pathophysiology, Guangdong Medical College, Dongguan 523808, China.
- Publication Type:Journal Article
- MeSH:
Carcinoma, Hepatocellular;
genetics;
pathology;
Cell Line, Tumor;
Gene Transfer Techniques;
Humans;
Liver Neoplasms;
genetics;
pathology;
Nanoparticles;
chemistry;
Polyethylene Glycols;
chemistry;
Polyethyleneimine;
chemistry;
Transfection;
methods
- From:
Journal of Biomedical Engineering
2011;28(1):104-109
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to develop non-toxic, high transfection efficiency polyethyleneimine(PEI) cationic nanoparticles. The exosyndrome of PEI cationic nanoparticles was measured by zeta sizer, ultraviolet and visible spectroscopy. The condensation ability and the resistance to DNaseI of pEGFP-N1/PEI and pEGFP-N1/PEI modified polyethylene glycol(PEG) were evaluated by agarose gel electrophoresis. The cell toxicity of polyethyleneimine cationic nanoparticles was measured by using MTT test. The orthogonal design was used to optimize the transfection efficiency with the N/P ratio, the grafting ratio and the gene dosage as the factors. The experimental results showed that pEGFP-N1/PEI nanoparticles have lower cell toxicity, better composite ability and better resistance to DNAseI. The highest transfection efficiency of PEI cationic nanoparticles was 91% by using the PEI nanoparticles with the N/P ratio 40:1 and gene dosages 6 microg/well. PEI cationic nanoparticle modified by PEG effectively transferred DNA to hepatoma carcinoma cells and it is a non-toxic, with high transfection efficiency, and a promising non-viral carrier for gene delivery. The transfection efficiency will be improved by optimizing the experiment condition.
