Transfection with a novel cationic gene carrier: PEI-PBLG.
- Author:
Ji-Zheng WEI
1
;
Lei LIN
;
Wei XIONG
;
Qing-Yu ZHU
Author Information
1. State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China.
- Publication Type:Journal Article
- MeSH:
Animals;
COS Cells;
Cell Line;
Cell Survival;
drug effects;
Cercopithecus aethiops;
DNA;
chemistry;
genetics;
ultrastructure;
Flow Cytometry;
Green Fluorescent Proteins;
genetics;
metabolism;
HeLa Cells;
Humans;
Microscopy, Electron, Scanning;
Microscopy, Fluorescence;
Molecular Weight;
Plasmids;
chemistry;
genetics;
Polyethyleneimine;
analogs & derivatives;
chemistry;
pharmacology;
Polyglutamic Acid;
analogs & derivatives;
chemistry;
pharmacology;
Transfection;
methods;
Vero Cells
- From:
Chinese Journal of Biotechnology
2007;23(2):229-234
- CountryChina
- Language:Chinese
-
Abstract:
This study mainly deals with cell transfection and cytotoxicity for PEI(10kD)-PBLG, a novel cationic copolymer, to observe its potential as a gene carrier. Size measurement and SEM were used to show the modality of the PEI-PBLG/pDNA complexes. Cytotoxicity of PEI (10kD)-PBLG was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay and compared with PEI(25kD)-PBLG, PEI(10kD), and PEI(25kD). Furthermore, pEGFP that can express the enhanced green fluorescent protein was chosen as a reporter to observe the transfection efficiency directly. Then, PEI (10kD)-PBLG/pEGFP complexes were transfected into several cell lines, such as Hela, COS-7, Vero-E6, and ECV-304, and effects of the transfection conditions were evaluated. The efficiencies were measured by FACS. Size measurement of complex particles indicated that PEI-PBLG/pDNA tended to form smaller nanoparticles compared with PEI/pDNA. The representative size of the PEI(10kD)-PBLG/pDNA complex was approximately 100 - 200 nm. SEM images showed that the particles were condense and compact. This can be suitable for their entry into cells. Cytotoxicity studies suggested that PEI (10kD)-PBLG had considerably lower toxicity than the other three materials. In the transfection tests, PEI (10kD)-PBLG/pDNA complexes could be transfected into all the cell lines that were tested. These provided the highest level of EGFP expression (45.02%) in Hela cells, which was considerably higher than that of PEI(10kD)/pEGFP (29.16%). Being less affected by the serum during transfection, PEI-PBLG/pDNA complexes offered greater biocompatibility than PEI. PEI-PBLG copolymer reduces the cytotoxicity of PEI, improves the transfection efficiency, and offers greater biocompatibility than PEI. It shows considerable potential as an efficient nonviral carrier for gene delivery.
