High efficiency transfection of primary skeletal muscle cells with lipid-based reagents.
Lipofection is a convenient method for gene transfer into muscle cells but reportedly is inefficient. We tested the efficacy of commercially available lipid-based and polyamine transfection reagents. Primary rat skeletal muscle cell cultures were transfected at three stages of development and assayed after fusion. Efficiency reached 30% during the proliferation stage and up to 23% when most myoblasts had fused into myotubes. Optimization of transfection conditions with three different vectors yielded efficiencies exceeding 50%. Thus, lipid-based transfection into primary skeletal muscle cells can be several times more efficient than previously reported.
Laboratory of Cell Biology, National Heart, Lung and Blood Institute, National Institutes of Health, 50 South Drive, Bethesda, Maryland 20892-8017, USA.
Birgit
B
Neuhuber
David I
DI
Huang
Mathew P
MP
Daniels
Carol E
CE
Torgan
D000818:Animals; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D005240:Feasibility Studies; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents / Q000494:pharmacology*; D008055:Lipids / Q000494:pharmacology*; D008164:Luminescent Proteins / Q000235:genetics; D018482:Muscle, Skeletal / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000235:genetics
C411955:FuGene; D007202:Indicators and Reagents; D008055:Lipids; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase; D001616:beta-Galactosidase