High efficiency transfection of primary skeletal muscle cells with lipid-based reagents

Muscle Nerve. 2002 Jul;26(1):136-40. doi: 10.1002/mus.10171.

Abstract

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.

MeSH terms

  • Animals
  • Cells, Cultured
  • Chloramphenicol O-Acetyltransferase / genetics
  • DNA / genetics
  • DNA / metabolism*
  • Feasibility Studies
  • Gene Expression / drug effects
  • Genes, Reporter
  • Genetic Vectors
  • Green Fluorescent Proteins
  • Indicators and Reagents / pharmacology*
  • Lipids / pharmacology*
  • Luminescent Proteins / genetics
  • Muscle, Skeletal / cytology
  • Muscle, Skeletal / drug effects*
  • Muscle, Skeletal / metabolism
  • Rats
  • Transfection / methods*
  • beta-Galactosidase / genetics

Substances

  • FuGene
  • Indicators and Reagents
  • Lipids
  • Luminescent Proteins
  • Green Fluorescent Proteins
  • DNA
  • Chloramphenicol O-Acetyltransferase
  • beta-Galactosidase