Nucleofection Induces Non-Specific Changes in the Metabolic Activity of Transfected Cells

Mol Biol Rep. 2012 Mar;39(3):2187-94. doi: 10.1007/s11033-011-0967-z. Epub 2011 Jun 5.


Transfection has become an everyday technique widely used for functional studies in living cells. The choice of the particular transfection method is usually determined by its efficiency and toxicity, and possible functional consequences specific to the method used are normally overlooked. We describe here that nucleofection, a method increasingly used because of its convenience and high efficiency, increases the metabolic rate of some cancer cells, which can be misleading when used as a measure of proliferation. Moreover, nucleofection can alter the subcellular expression pattern of the transfected protein. These undesired effects are independent of the transfected nucleic acid, but depend on the particular cell line used. Therefore, the interpretation of functional data using this technology requires further controls and caution.

MeSH terms

  • Analysis of Variance
  • Animals
  • Cell Line
  • Cell Nucleus / metabolism*
  • Cell Proliferation
  • Electroporation / methods*
  • Energy Metabolism / physiology*
  • Flow Cytometry
  • Humans
  • Mice
  • Microscopy, Fluorescence
  • Oxazines / metabolism
  • Oxidation-Reduction
  • Real-Time Polymerase Chain Reaction
  • Tetrazolium Salts / metabolism
  • Thiazoles / metabolism
  • Transfection / methods*
  • Xanthenes / metabolism


  • Oxazines
  • Tetrazolium Salts
  • Thiazoles
  • Xanthenes
  • resazurin
  • thiazolyl blue