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Adenoviral transduction is more efficient in alginate-derived chondrocytes than in monolayer chondrocytes

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Abstract

Gene transfer into cultured chondrocytes by using adenoviral vectors has potential applications in treating cartilage disorders. The present study was undertaken to compare and optimize two chondrocyte culture conditions for adenoviral transduction efficacy by using primary human articular chondrocytes cultivated either directly in a monolayer condition or as outgrowths from alginate-stored chondrocyte cultures. Isolated primary chondrocytes from human articular cartilage were either immediately transduced with an EGFP (enhanced green fluorescent protein)-gene-bearing adenoviral vector (1,000 and 3,000 virus particles/cell) or cultured in alginate before transduction. Immunohistochemistry and flow cytometric analysis were employed to determine the expression of extracellular matrix proteins and of the αvβ5 integrin receptor involved in adenoviral cell entry. Monolayer chondrocytes exhibited moderate transduction rates (mean 22.2% and 46.9% EGFP-positive cells at 1,000 and 3,000 virus particles/cell by 72 h post-transduction), whereas alginate-derived chondrocytes revealed significantly higher transduction efficacies (95.7% and 99%). Both monolayer and alginate-derived chondrocytes expressed αvβ5 integrin, type II collagen and cartilage proteoglycans. The mean fluorescence intensity of type II collagen was significantly higher in the alginate-derived chondrocytes, whereas that of αvβ5 integrin was higher in the monolayer chondrocytes. Our results indicate that transduction efficacy is independent of αvβ5 integrin expression levels in chondrocytes. Moreover, adenoviral transduction of alginate-derived chondrocytes is more efficient than that for monolayer chondrocytes and may be a suitable tool to achieve sufficient numbers of transduced and differentiated chondrocytes for experimental applications and cartilage repair.

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Acknowledgement

The authors are indebted to Mrs. Marion Lemke for her excellent technical assistance.

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Authors and Affiliations

  1. Department of Trauma and Reconstructive Surgery, Charité Medical Schools Berlin, Campus Benjamin Franklin, FEM, Krahmerstrasse 6-10, 12207, Berlin, Germany

    A. Oberholzer, T. John, B. Kohl, T. Gust, R. D. Müller, W. Ertel & G. Schulze-Tanzil

  2. Centrum of Anatomy, Department of Cell and Neurobiology, Charité Medical Schools Berlin, Campus Benjamin Franklin, Berlin, Germany

    G. Schulze-Tanzil

  3. Biomaterials & Tissue Engineering Research Unit, Biomedical Engineering, School of AMMEJ07, University of Sydney, Sydney, NSW, 2006, Australia

    H. Zreiqat

  4. Canji, San Diego, CA, USA

    D. La Face & B. Hutchins

Authors
  1. A. Oberholzer
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  2. T. John
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  3. B. Kohl
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  4. T. Gust
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  5. R. D. Müller
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  6. D. La Face
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  7. B. Hutchins
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  8. H. Zreiqat
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  9. W. Ertel
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  10. G. Schulze-Tanzil
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Corresponding author

Correspondence to G. Schulze-Tanzil.

Additional information

Dr. Gundula Schulze-Tanzil is supported by a grant awarded by the Rahel Hirsh Foundation from the Charité Medical Schools Berlin. The study was supported by a grant from the Deutsche Arthrosehilfe e.V.

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Oberholzer, A., John, T., Kohl, B. et al. Adenoviral transduction is more efficient in alginate-derived chondrocytes than in monolayer chondrocytes. Cell Tissue Res 328, 383–390 (2007). https://doi.org/10.1007/s00441-006-0371-5

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  • DOI: https://doi.org/10.1007/s00441-006-0371-5

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