DOTAP/DOPE ratio and cell type determine transfection efficiency with DOTAP-liposomes
DOTAP/DOPE ratio and cell type determine transfection efficiency with DOTAP-liposomes
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DOI：
10.1016/j.bbamem.2015.06.020
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发表时间：
2015-10-01
影响因子：
3.4
通讯作者：
Doh, Kyung-Oh
中科院分区：
文献类型：
--
作者：
Kim, Bieong-Kil;Hwang, Guen-Bae;Doh, Kyung-Oh
研究了脂质组成对其理化性质和转染效率的影响。研究了具有不同1，2-二油酰基-3-三甲基铵丙烷（DOTAP）与二油酰基磷脂酰乙醇胺（DOPE）重量比的四种脂质体制剂，即重量比1：0（T1 P0）、3：1（T3 P1）、1：1（T1 P1）和1：3（T1 P3）。脂质体的平均大小受其脂质组成和超声处理时的制剂浓度的影响。脂质体的Zeta电位与其脂质体的大小呈负相关。然而，无论是脂质体的大小，也没有zeta电位与转染效率。脂质体的最佳组成是细胞系依赖性的（对于Huh 7和AGS，TIP和T3 P1;对于COS 7，T3 P1和T1 P1;对于A549，T1 P1和T1 P3）。脂质复合物的形状根据DOPE比例的增加而从层状结构变为倒置六边形结构，但在所有使用的细胞系中，特定结构在转染效率上没有明确的优势。然而，在所有细胞系中，与T1 P3相比，TIP、T3 P1、T1 P1中的细胞内化始终更快，这表明内体逃逸的重要性。我们的研究结果表明，DOTAP脂质体的转染效率主要受脂质体组成和细胞类型的影响，而不是由大小或zeta电位。(C)2015 Elsevier B. V.版权所有。
The effects of lipid compositions on their physicochemical properties and transfection efficiencies were investigated. Four liposome formulations with different 1,2-dioleoy1-3-trimethylammoniumpropane (DOTAP) to dioleoylphosphatidylethanolamine (DOPE) weight ratios were investigated, that is, weight ratios 1:0 (T1P0), 3:1 (T3P1), 1:1 (T1P1), and 1:3 (T1P3). Mean sizes of liposomes were influenced by their lipid composition and the preparation concentration at the time of sonication. Zeta potentials of liposomes were inversely correlated with their liposome sizes. However, neither liposome sizes nor zeta potentials were correlated with transfection efficiency. The optimum composition of liposomes was cell-line dependent (TIP and T3P1 for Huh7 and AGS, T3P1 and T1P1 for COS7, and T1P1 and T1P3 for A549). The shape of lipoplexes was changed from lamellar to inverted hexagonal structure according to the increased ratio of DOPE, but there was no definite advantage of specific structure in transfection efficiency throughout all used cell lines. However, cellular internalization was consistently faster in TIP, T3P1, T1P1 compared to T1P3 in all cell lines, suggesting the importance of endosomal escape. Our findings show that the transfection efficiency of DOTAP liposomes is mainly influenced by lipid composition and cell type, and not by size or zeta potential. (C) 2015 Elsevier B.V. All rights reserved.