Comparative evaluation of gene delivery devices in primary cultures of rat hepatic stellate cells and rat myofibroblasts

With the use of chemically enhanced transfection methods, the highest relative efficiency was obtained with FuGENE？6 gene mediated DNA transfer. Quantitative evaluation of representative transfection experiments by flow cytometry revealed that approximately 6% of the rat hepatic stellate cells were transfected. None of the transfection methods tested was able to mediate gene delivery to rat myofibroblasts. To analyze if rat hepatic stellate cells and myofibroblasts are susceptible to adenoviral infection, we have inserted the transgenic expression cassette into a recombinant adenoviral type 5 genome as replacement for the E1 region. Viral particles of this replication-deficient Ad5-based reporter are able to infect 100% of rat hepatic stellate cells and myofibroblasts, respectively.Our results indicate that FuGENE？6-based methods may be optimized sufficiently to offer a feasible approach for gene transfer into rat hepatic stellate cells. The data further demonstrate that adenoviral mediated transfer is a promising approach for gene delivery to these hepatic cells.Transfection is the insertion of foreign molecules such as cDNAs or promoter constructs into eukaryotic cells. This method has become a powerful experimental tool for studying gene functions and to analyze the control of gene expression. Genes of interest can either be transfected transiently or stable into cultured mammalian cells. Detailed protocols for efficient gene transfer to various primary cells and continuous cell lines, irrespective whether these cells are grown as monolayers or in suspension have been established during the last decades. Many methods have been developed to overcome the low transfection efficiency in differentiated cells if classical approaches, such as calcium-phosphate-DNA coprecipitates [1], diethylaminoethyl (DEAE)-dextran [2] or polylysine mediated gene transfer [3] are applied. Transfection by electroporation, microinjection, biolistic particle delivery, activated dendrimers