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The Journal of Membrane Biology

, Volume 248, Issue 2, pp 273–283 | Cite as

Electrotransfection and Lipofection Show Comparable Efficiency for In Vitro Gene Delivery of Primary Human Myoblasts

  • Tomaz MarsEmail author
  • Marusa Strazisar
  • Katarina Mis
  • Nejc Kotnik
  • Katarina Pegan
  • Jasna Lojk
  • Zoran Grubic
  • Mojca PavlinEmail author
Article

Abstract

Transfection of primary human myoblasts offers the possibility to study mechanisms that are important for muscle regeneration and gene therapy of muscle disease. Cultured human myoblasts were selected here because muscle cells still proliferate at this developmental stage, which might have several advantages in gene therapy. Gene therapy is one of the most sought-after tools in modern medicine. Its progress is, however, limited due to the lack of suitable gene transfer techniques. To obtain better insight into the transfection potential of the presently used techniques, two non-viral transfection methods—lipofection and electroporation—were compared. The parameters that can influence transfection efficiency and cell viability were systematically approached and compared. Cultured myoblasts were transfected with the pEGFP-N1 plasmid either using Lipofectamine 2000 or with electroporation. Various combinations for the preparation of the lipoplexes and the electroporation media, and for the pulsing protocols, were tested and compared. Transfection efficiency and cell viability were inversely proportional for both approaches. The appropriate ratio of Lipofectamine and plasmid DNA provides optimal conditions for lipofection, while for electroporation, RPMI medium and a pulsing protocol using eight pulses of 2 ms at E = 0.8 kV/cm proved to be the optimal combination. The transfection efficiencies for the optimal lipofection and optimal electrotransfection protocols were similar (32 vs. 32.5 %, respectively). Both of these methods are effective for transfection of primary human myoblasts; however, electroporation might be advantageous for in vivo application to skeletal muscle.

Keywords

Myoblasts Lipofection Electroporation Gene electrotransfer Gene therapy 

Notes

Acknowledgments

This study was supported by the Slovenian Research Agency (Grant No. J4-4324, research programme P3-0043). We gratefully acknowledge help from Zvonka Frelih, Marko Narobe and Romain Teruel.

Conflict of interest

The authors report no conflicts of interest.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Tomaz Mars
    • 1
    Email author
  • Marusa Strazisar
    • 2
  • Katarina Mis
    • 1
  • Nejc Kotnik
    • 1
  • Katarina Pegan
    • 1
  • Jasna Lojk
    • 2
  • Zoran Grubic
    • 1
  • Mojca Pavlin
    • 2
    Email author
  1. 1.Institute of Pathophysiology, Faculty of MedicineUniversity of LjubljanaLjubljanaSlovenia
  2. 2.Faculty of Electrical EngineeringUniversity of LjubljanaLjubljanaSlovenia

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