Nejc Kotnik¹, Saša Haberl², Katarina Pegan¹, Zoran Grubič¹, Mojca Pavlin², Tomaž Marš¹
¹Institute of Pathophysiology, Faculty of Medicine, University of Ljubljana, Slovenia
²Faculty of Electrical Engineering, University of Ljubljana, Slovenia
One of main goals of in vitro research is to develop efficient methods for drug delivery. Effective delivery of genetic material into cells is crucial for application in clinical environment for gene therapy or genetic vaccination. The tissue of choice for gene therapy or DNA vaccination are skeletal muscles, representing 40 % of the body mass and which are known to actively participate in the immune response.
Here we compared the efficiency of two different methods for transfer of plasmid DNA: electroporation, where electric pulses are used to permeabilize cell membrane; and lipofection which uses liposomes as carriers. Electroporation is promising for clinical application while lipofection is auspicious for developing strategies for gene therapy. Plasmid pEGFP-N1 coding for green fluorescent protein was transfected into primary human myoblasts with lipofection (Lipofectamine 2000) or by applying high-voltage pulses.
Our results demonstrate that efficient transfection of human myoblasts with DNA in vitro can be obtained both methods. The highest transfection ratios were similar (40.9% with lipofection versus 41.4% with electroporation). However the main difference was in cytotoxicity of both methods. Less than 40% of cells were viable after the electroporation under conditions that give comparable transfection results with lipofection. On the other hand, by lipofection, almost 80% of myoblast were viable after the treatment. Due to lower cytotoxicity, more suitable method for transfection studies on myoblasts in vitro is lipofection. Electroporation has advantage over lipofection in in vivo environment due to simplicity of protocol and lack of need for using additional chemicals.
Keywords: myoblast, lipofection, electroporation, gene therapy