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題 名 | Transfection of BHK Cell by Serum-Stabilized Cationic Liposome-DNA Particles |
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作 者 | Huang, Yi-you; Cullis, Pieter R; | 書刊名 | 醫學工程 |
卷 期 | 12:6 民89.12 |
頁 次 | 頁281-287 |
分類號 | 410.013366 |
關鍵詞 | 基因轉殖; Cationic liposome; Transfection; BHK cell; Serum-stabilized liposome; |
語 文 | 英文(English) |
英文摘要 | Cationic liposomes complexed with DNA have been used extensively as non-viral vectors for the intracellular delivery of reporter or therapeutic genes in cell culture can vivo transfection experiments. Most of cationic liposome-DNA particles will be cleared from the blood very quickly when they were administered into the blood circulation system. Serum-stabilized cationic liposome-DNA particles made by preformed vesicles and ethanol method were developed [1]. Steric stabilization cofers long circulation time to these particles, allowing them to extravasate more easily at sites of porous vasculature. In vitro transfection potency was evaluated by culturing with BHK cells. Experimental results show that the cell uptake of cationic liposome-DNA particles made by DO-DAP/DSPC/Chol/PEG-CerC14 (25/20/45/10 mol%) was higher than that of made by DO-DAP/DOPE/Chol/PEG-CerC14 (20/50/20/10 mol%). However, the transfection efficiency of liposome-DNA particles made by DODAP/DOPE/Chol/PEG-CerC14 (20/50/20/10 mol%) was much higher than the liposome-DNA particles made by DODAP/DSPC/Chol/PEG-CerC14 (25/20/45/10 mol%). This confirms that DOPE is a transfection helper lipid. Except the COPE, the concentration of calcium ion also plays an important role in the BHK transfection experiments. 10 mM Ca++ was necessary for achieving high transfection efficiency. |
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