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  • Nonviral Transfer Technology
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Efficient lipid-mediated gene transfer to articular chondrocytes

Gene Therapy volume 7, pages 286–291 (2000)Cite this article

Abstract

We examined nonviral, lipid-mediated gene transfer methods as potential tools for efficient transfection of articular chondrocytes. Transfection conditions were determined for primary cultures of normal human articular, osteoarthritic human articular and normal bovine articular chondrocytes using a lacZ reporter gene construct with the commercially available cationic liposomes Cellfectin, DMRIE-C, LipofectAmine, Lipofectin, LipoTaxi, TransFast and the lipid-based reagent FuGENE 6. Optimized conditions were then evaluated in an ex vivo model of chondrocyte transplantation. FuGENE 6 transfection produced the maximum levels of transgene expression. Transfection efficiency was cell type specific and affected by DNA concentration, lipid/DNA ratio and the presence of hyaluronidase, a matrix-degrading enzyme. Analysis of X-gal staining demonstrated an efficiency of 41.0% in normal bovine articular chondrocytes, 20.7% in normal human articular chondrocytes and 7.8% in osteoarthritic human chondrocytes. Transfected chondrocytes were found to successfully populate the articular cartilage surface in explant cultures. Transplanted genetically modified chondrocytes adhered to the articular cartilage and continued to produce β-galactosidase for 2 weeks. This evaluation and optimization of lipid-based gene transfer into articular chondrocytes may serve as a useful tool in studies of genes involved in articular cartilage damage and repair and as a potential delivery method for therapeutic genes.

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Acknowledgements

We thank T Fondren, H Mankin and D Burke for human articular cartilage specimens, C Trahan for histological processing, L Weissbach and M Cucchiarini for helpful discussions. This work was supported by the German Academy for Natural Scientists LEOPOLDINA and NIH grants AR31068 and AR45749-01.

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  1. Department of Orthopaedic Surgery, Orthopaedic Research Laboratories, Harvard Medical School and Massachusetts General Hospital, Boston, MA, USA

    H Madry & S B Trippel

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  1. H Madry
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Madry, H., Trippel, S. Efficient lipid-mediated gene transfer to articular chondrocytes. Gene Ther 7, 286–291 (2000). https://doi.org/10.1038/sj.gt.3301086

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