Skip to main content

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 229))

  • 1146 Accesses

Abstract

A mammalian cornea can be maintained in vitro for days (1,2), allowing ex vivo transduction of this tissue prior to transplantation (3,4). Ex vivo transduction of the cornea is useful for studying the efficacy of gene products on inhibition of corneal neovascularization (4), amelioration of corneal inflammatory disease, or promotion of corneal wound healing in animal models. Ex vivo transduction can also be used for transfer of genes into corneal epithelial stem cells (5), thereby allowing genetically modified stem cells to be reimplanted into the limbal area (6), providing a means for long-term gene-replacement (e.g., βig-h3 gene) therapy for various inherited corneal diseases (7).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 74.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Frueh, B. E. and Bohnke, M. (1995) Corneal grafting of donor tissue preserved for longer than 4 weeks in organ-culture medium. Cornea 14, 463–466.

    Article  PubMed  CAS  Google Scholar 

  2. Redbrake, C., Sallas S., and Frantz, A. (1998) Changes in human donor corneas preserved for longer than 4 weeks. Cornea 17, 62–65.

    Article  PubMed  CAS  Google Scholar 

  3. Larkin, D. F., Oral, H. B., Ring, C. J., Lemoine, N. R., and George, A. J. (1996) Adenovirus-mediated gene delivery to the corneal endothelium. Transplantation 61, 363–370.

    Article  PubMed  CAS  Google Scholar 

  4. Yoken, J., Chen, S. L., McFarland, T., et al. (2002) Production of a lentiviral vector containing an endostatin:kringle-5 fusion gene an inhibitor of post-penetrating keratoplasty neovascularization. IOVS Presentation S1868.

    Google Scholar 

  5. Bradshaw, J. J., Obritsch, W. F., Cho, B. J., Gregerson, D. S., and Holland, E. J. (1999) Ex vivo transduction of corneal epithelial progenitor cells using a retroviral vector. IOVS 40, 230–235.

    CAS  Google Scholar 

  6. Tsubota, K., Satake, Y., Kaido, M., et al. (1999) Treatment of severe ocular-surface disorders with corneal epithelial stem-cell transplantation. N. Eng. J. Med. 340, 1697–1703.

    Article  CAS  Google Scholar 

  7. Munier, F. L., Korvatska, E., Djemai, A., et al. (1997) Kerato-epithelin mutations in four 5q31-linked corneal dystrophies. Nat. Gen. 15, 247–251.

    Article  CAS  Google Scholar 

  8. Price, F. W., Whitson, W. E., Collins, K. S., and Marks R. G. (1993) Five-year corneal graft survival. Arch. Ophthalmol. 111, 799–805.

    PubMed  Google Scholar 

  9. Vail, A., Gore, S. M., Bradley, B. A., Easty, D. L., and Rogers, C. A. (1993) Corneal transplantation in the United Kingdom and Republic of Ireland. Br. J. Ophthalmol. 77, 650–652.

    Article  PubMed  CAS  Google Scholar 

  10. Bourne W. M., Hodge D. O., and Nelson L. R. (1994) Corneal endothelium five years after transplantation. Am. J. Ophthalmol. 118, 185–196.

    PubMed  CAS  Google Scholar 

  11. Oral, H. B., Larkin, D. F., Fehervari, Z., et al. (1997) Ex vivo adenovirus-mediated gene transfer and immunomodulatory protein production in human cornea. Gene Ther. 4, 639–647.

    Article  PubMed  CAS  Google Scholar 

  12. Fehervari, Z., Rayner, S. A., Oral, H. B., George, A. J., and Larkin, D. F. (1997) Gene transfer to ex vivo stored corneas. Cornea 16, 459–464.

    Article  PubMed  CAS  Google Scholar 

  13. Budenz, D. L., Bennett, J., Alonso, L., and Maguire, A. (1995) In vivo gene transfer into murine corneal endothelial and trabecular meshwork cells. IOVS 36, 2211–2215.

    CAS  Google Scholar 

  14. Tanelian, D. L., Barry, M. A., Johnston, S. A., Le, T., and Smith, G. (1997) Controlled gene gun delivery and expression of DNA within the cornea. Biotechniques 23, 484–488.

    PubMed  CAS  Google Scholar 

  15. Oshima, Y., Sakamoto, T., Yamanaka, I., Nishi, T., Ishibashi, T., and Inomata, H. (1998) Targeted gene transfer to corneal endothelium in vivo by electric pulse. Gene Ther. 5, 1347–1354.

    Article  PubMed  CAS  Google Scholar 

  16. Wang, X., Appukuttan, B., Ott, S., et al. (2000) Efficient and sustained transgene expression in human corneal cells mediated by a lentiviral vector. Gene Ther. 7, 196–200.

    Article  PubMed  CAS  Google Scholar 

  17. Yang, X.-J. (2001) Preparation of recombinant retroviruses, in Vision Research Protocols (Rakoczy, P. E., ed.), Humana, Totowa, NJ, pp. 171–190.

    Google Scholar 

  18. Gorman, C. M., Howard, B. H., and Reeves, R. (1983) Expression of recombinant plasmids in mammalian cells is enhanced by sodium butyrate. Nucleic Acids Res. 11, 7631–7648.

    Article  PubMed  CAS  Google Scholar 

  19. Murthy, R. C., McFarland, T. J., Yoken, J., et al. (2003) Ex vivo transduction of corneas with a novel fusion gene inhibits angiogenesis and graft failure in a corneal transplantation model. Invest. Opthalmol. Vis. Sci., in press.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2003 Humana Press Inc.

About this protocol

Cite this protocol

Appukuttan, B., McFarland, T., Stout, J.T. (2003). Corneal Cells. In: Federico, M. (eds) Lentivirus Gene Engineering Protocols. Methods in Molecular Biology™, vol 229. Humana Press. https://doi.org/10.1385/1-59259-393-3:197

Download citation

  • DOI: https://doi.org/10.1385/1-59259-393-3:197

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-091-5

  • Online ISBN: 978-1-59259-393-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics