Transfection with a novel cationic gene carrier: PEI-PBLG]

Chinese Journal of Biotechnology(2007)

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摘要
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 performed 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 facial action coding system (FACS). Size measurement of complex particles indicated that PEI-PBLG/pDNA tended to form smaller nanoparticles compared with PEI/pDNA. The typical size of the PEI(10kD)-PBLG/pDNA complex was approximately 100–200 nm. SEM images showed that the particles were condensed 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.
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关键词
polyethylenimine (PEI),poly(β-benzyl L-glutamate) (PBLG),gene delivery,transfection efficiency,cytotoxicity
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