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  • 标题:Comparison of the Efficiency and Safety of Non-viral Vector-Mediated Gene Transfer into a Wide Range of Human Cells
  • 本地全文:下载
  • 作者:Eriko Uchida ; Hiroyuki Mizuguchi ; Akiko Ishii-Watabe
  • 期刊名称:Biological and Pharmaceutical Bulletin
  • 印刷版ISSN:0918-6158
  • 电子版ISSN:1347-5215
  • 出版年度:2002
  • 卷号:25
  • 期号:7
  • 页码:891-897
  • DOI:10.1248/bpb.25.891
  • 出版社:The Pharmaceutical Society of Japan
  • 摘要:Non-viral gene transfer into a wide range of human cells was examined in order to clarify the factors that affect the efficiency and safety of non-viral vectors and to optimize the conditions so that high efficiency and low toxicity could be achieved. Six non-viral vectors (Lipofectin, LipofectAMINE PLUS, SuperFect, Effectene, DMRIE-C and DOTAP) were used to transfect a mammalian expression plasmid pCMVβ into 16 types of human primary cells and cultured cell lines. Transfection efficiency was quantified using a galactosidase assay. Cytotoxic effects were measured by lactate dehydrogenase (LDH) assay and WST-8 assay. In serum-free conditions, LipofectAMINE PLUS, Effectene and SuperFect, on average, transfected DNA more successfully than Lipofectin, DMRIE-C, and DOTAP, although the levels of gene expression with these vectors varied remarkably in different cells. The most effective vector also differed depending on the cell type. Serum was found to inhibit gene transfer and reduce the cytotoxicity of all of these vectors except Effectene. The efficiency and toxicity of the non-viral vectors used depended on the type of vector, the DNA/vector ratio, the type of cell, and the presence of serum. These results provided useful information for the optimization of transfer conditions of these non-viral vectors.
  • 关键词:gene transfer;non-viral vector;transfection efficiency;cytotoxicity
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