Use of electroporation to introduce biologically active foreign genes into primary rat hepatocytes.
A method is described for introducing and expressing cloned genes in isolated hepatocytes. Primary rat hepatocytes isolated by collagenase perfusion were transfected in suspension with plasmid pSV2CAT by electroporation. Forty-eight hours later, soluble extracts from transfected hepatocytes showed chloramphenicol acetyltransferase activity comparable to that obtained in rat hepatoma cell line H4AzC2 by calcium phosphate or DEAE-dextran transfection. The latter two methods could not be used successfully for primary hepatocytes because of cytotoxicity of these reagents. This indicates that electroporation is a useful method to obtain transient expression of foreign genes in primary epithelial cells, such as rat hepatocytes, which are difficult to maintain in cell culture.
Tur-Kaspa
R
R
Teicher
L
L
Levine
B J
BJ
Skoultchi
A I
AI
Shafritz
D A
DA
D000123:Acetyltransferases / Q000235:genetics; D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase; D003001:Cloning, Molecular; D004566:Electrodes; D005796:Genes; D008099:Liver / Q000378:metabolism*; D008114:Liver Neoplasms, Experimental; D010957:Plasmids; D051381:Rats
D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase