Culture time-dependent gene expression in isolated primary cultured rat hepatocytes by transfection with the cationic liposomal vector TFL-3.
Culture time-dependent gene expression in isolated primary cultured rat hepatocytes by transfection with the cationic liposomal vector TFL-3.
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通过用阳离子脂质体载体 TFL-3 转染，在分离的原代培养大鼠肝细胞中培养时间依赖性基因表达。
DOI：
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发表时间：
2003
影响因子：
2
通讯作者：
H. Kiwada
中科院分区：
文献类型：
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作者：
Lap Thi Nguyen;T. Ishida;Sachiko Ukitsu;W. Li;R. Tachibana;H. Kiwada
开发能够将功能性外源基因转移到不分裂或分裂频率较低的哺乳动物细胞的载体系统对于增加治疗各种疾病的可用选择是必不可少的。TFL-3，最近开发的阳离子脂质体，是否可以成功地用于实现在原代培养的大鼠肝细胞中的基因表达的问题进行了检查。在4小时预培养后的不同时间点，用TFL-3中的质粒DNA（pDNA）转染肝细胞4小时。在转染后的不同时间用编码氯霉素乙酰转移酶（CAT）、荧光素酶或β-半乳糖苷酶的pDNA测定转染效率。作为脂质转染的结果，还定量测定了存在的核内pDNA的量。在我们的实验条件下，对于所有测试的pDNA观察到成功的脂质转染，并且效率上级市售的LIPOFECTAMINE。基因表达的程度和速率取决于脂质转染前的孵育时间以及每皿细胞的密度，但这种关系并不适用于传递到细胞核的基因量。这些结果表明，TFL-3可能是一个有用的载体，为实现足够的基因表达在大鼠肝细胞，并建议，培养时间之前和之后的脂质体转染，这是相关的细胞的生物学条件，可能是一个主要因素，影响有效的基因表达在非分裂细胞。
The development of a carrier system that enables the transfer of a functional exogenous gene to non- or less frequently dividing mammalian cells is essential for increasing the available options for the treatment of various diseases. The issue of whether TFL-3, a recently developed cationic liposome, can be successfully used to achieve gene expression in primary cultured rat hepatocytes was examined. The hepatocytes were transfected for 4 h with plasmid DNA (pDNA) in TFL-3 at various time points after 4-h preculture. The transfection efficiency was determined at various times posttransfection with pDNA coding for chloramphenicol acetyltransferase (CAT), luciferase, or beta-galactosidase. The amount of intranuclear pDNA present, as a consequence of the lipofection, was also quantitatively determined. Successful lipofections were observed for all pDNA tested, and the efficiencies were superior to that of commercially available LIPOFECTAMINE under our experimental conditions. The degree and rate of gene expression were dependent on incubation time prior to lipofection as well as on the density of the cells per dish, but this relationship did not hold for the amount of gene delivered to the nuclei. These results indicate that TFL-3 could be a useful vector for achieving sufficient gene expression in rat hepatocytes and suggest that the culture time prior to and following lipofection, which is related to the biological condition of the cells, may be one major factor affecting efficient gene expression in nondividing cells.
影响因子：
56.9
作者：
RHIM, JA;SANDGREN, EP;BRINSTER, RL
通讯作者：
BRINSTER, RL
影响因子：
4.2
作者：
H. Temin
通讯作者：
H. Temin