Transfection of primary cultures of rat hepatocytes.
Five different transfection reagents-calcium phosphate, TransFast Transfection Reagent, Superfect Transfection Reagent, Effectene Transfection Reagent, and Tfx-20--were compared for their ability to effectively transfect primary cultures of male rat hepatocytes. Hepatocytes were isolated by the collagenase perfusion method and then cultured on Matrigel-coated plates for 24 h before transfection. The cells were transfected with either pGL3-Control or pGL3-Basic plasmids. The efficiency of transfection of each reagent was monitored using the dual luciferase reporter gene assay system. Superfect Transfection Reagent, Effectene Transfection Reagent and Tfx-20 were the most effective for the transfection of primary hepatocytes and gave comparable transfection efficiencies. Calcium phosphate was found to be the least effective transfection reagent and gave the most variable transfection results. Tfx-20 gave the least variable transfection results when different hepatocyte preparations were compared.
Edwards
Mina
M
Department of Biochemistry and Molecular Biology, University College London, UK.
Wong
Siew Cheng
SC
Chotpadiwetkul
Roongsiri
R
Smirlis
Despina
D
Phillips
lan R
lR
Shephard
Elizabeth A
EA
D000818:Animals; D002130:Calcium Phosphates; D002478:Cells, Cultured; D017930:Genes, Reporter; D022781:Hepatocytes / Q000166:cytology*; D008156:Luciferases / Q000235:genetics; D051381:Rats; D014162:Transfection
D002130:Calcium Phosphates; C020243:calcium phosphate; D008156:Luciferases