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.
Neuhuber
Birgit
B
Laboratory of Cell Biology, National Heart, Lung and Blood Institute, National Institutes of Health, 50 South Drive, Bethesda, Maryland 20892-8017, USA.
Huang
David I
DI
Daniels
Mathew P
MP
Torgan
Carol E
CE
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