Transfection of primary human skin fibroblasts by electroporation.
Primary human skin fibroblasts are an accessible source of phenotypically and karyotypically normal human cells, but are difficult to transfect with exogenous DNA. Here we demonstrate that both transient expression and stable transformation can be carried out by the method of electroporation. Highly efficient transient chloramphenicol acetyltransferase expression was shown after transfection with plasmid pRSVCAT. Stable transformation of human skin fibroblasts to G418 resistance was obtained after electroporation with neo-containing plasmids at an efficiency of approximately 1.4 x 10(-5)/micrograms DNA. The ability to easily transfect these cells with exogenous DNA may have important applications in the study of human genetic diseases and cancer.
Fountain
J W
JW
Howard Hughes Medical Institute, University of Michigan, Ann Arbor 48109.
Lockwood
W K
WK
Collins
F S
FS
D000328:Adult; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004560:Electricity; D005347:Fibroblasts / Q000378:metabolism; D005796:Genes; D005798:Genes, Bacterial; D006801:Humans; D010957:Plasmids; D012867:Skin / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
D015500:Chloramphenicol O-Acetyltransferase; D004262:DNA Restriction Enzymes