Comparison of long-term transgene expression after non-viral and adenoviral gene transfer into primary articular chondrocytes

Histochem Cell Biol. 2001 Jul;116(1):69-77. doi: 10.1007/s004180100305.


Different gene transfer approaches to achieve long-term transgene expression in cultured primary bovine chondrocytes were compared using enhanced green fluorescent protein (EGFP) as a reporter. Transduction with a high-capacity adenoviral vector was 82% efficient when analysed by fluorescence microscopy, while up to 42% of plasmid-transfected cells were EGFP positive with FuGene as a transfection reagent. Rapid dominant marker selection of plasmid-transfected cells was achieved in monolayer culture. With either method of gene transfer, a high proportion of the chondrocytes remained transgene positive during prolonged alginate culture. Transgene transcription in single cells was quantified with a confocal laser scanning microscope. Detection of EGFP expression was more sensitive with this method, identifying more transgene-expressing cells than conventional fluorescence microscopy. Long-term EGFP expression was higher in adenovirally transduced chondrocytes embedded in alginate as compared to plasmid-transfected cells cultured in monolayer or in alginate. Both the adenoviral and the plasmid-based approach appear suited for studies of the molecular and cellular mechanisms by which mutations in cartilage matrix proteins cause disease.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics*
  • Animals
  • Cattle
  • Chondrocytes / cytology
  • Drug Resistance, Microbial
  • Gene Expression / genetics*
  • Gene Transfer Techniques*
  • Genes, Reporter / genetics
  • Genetic Vectors / genetics
  • Green Fluorescent Proteins
  • Luminescent Proteins / analysis
  • Luminescent Proteins / genetics*
  • Microscopy, Confocal / instrumentation
  • Plasmids / genetics*
  • Transgenes / genetics


  • Luminescent Proteins
  • Green Fluorescent Proteins