Culture time-dependent gene expression in isolated primary cultured rat hepatocytes by transfection with the cationic liposomal vector TFL-3.

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.

Quantitative analysis of correlation between number of nuclear plasmids and gene expression activity after transfection with cationic liposomes.

Purpose: A quantitative understanding of the intracellular trafficking of plasmids delivered by nonviral vectors is essential for optimizing vector functions to increase their transfection efficiency. In this study, quantitative methods were developed to measure plasmids delivered to the nucleus, and the relationship between transfection activity and the number of plasmids in the nucleus were analyzed.

Methods: AH130 cells were transfected with plasmids in cationic liposomes at various doses.