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Corneal Assay for Angiogenesis

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Angiogenesis Protocols

Part of the book series: Methods in Molecular Medicineā„¢ ((MIMM,volume 46))

Abstract

In order to develop angiogenic and antiangiogenic strategies, concerted efforts have been made to provide animal models for more quantitative analysis of in vivo angiogenesis. In vivo techniques consist of the cornea pocket and iris implant in the eye, the rabbit ear chamber, the dorsal skinfold chamber, the cranial window, the hamster cheek pouch window, the sponge implant assay, the fibrin clots, the sodium alginate beads and the Matrigel plugs, the rat mesenteric window, the chick embryo chorioallantoic membrane and the air sac in mice and rats (1). In this chapter we will discuss the avascular cornea assay, and the advantages and disadvantages of using this assay in different species. The cornea assay is based on the placement of an angiogenic inducer (tumor tissue, cell suspension, growth factor) into a corneal pocket in order to evoke vascular outgrowth from the peripherally located limbal vasculature. In comparison to other in vivo assays, this assay has the advantage of measuring only new blood vessels, because the cornea is initially avascular.

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References

  1. Jain, R. K., Schlenger, K., Hockel, M., and Yuan, F. (1997) Quantitative angiogenesis assays: Progress and problems. Nature Med. 3, 1203ā€“1208.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  2. Gimbrone, M. Jr., Cotran, R., Leapman, S. B., and Folkman, J. (1974) Tumor growth and neovascularization: An experimental model using the rabbit cornea. J. Natl. Cancer Inst. 52, 413ā€“427.

    PubMedĀ  Google ScholarĀ 

  3. Ziche, M., Jones, J., and Gullino, P. M. (1982) Role of prostaglandin E1 and copper in angiogenesis.J. Natl. Cancer Inst. 69, 475ā€“482.

    CASĀ  PubMedĀ  Google ScholarĀ 

  4. Gallo, O., Masini, E., Morbidelli, L., Franchi, A., Fini-Storchi, I., Vergari, W. A., and Ziche, M. (1998) Role of nitric oxide in angiogenesis and tumor progression in head and neck cancer. J. Natl. Cancer Inst. 90, 587ā€“596.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  5. Langer, R. and Folkman, J. (1976) Polymers for the sustained release of proteins and other macromolecules. Nature 363, 797ā€“800.

    ArticleĀ  Google ScholarĀ 

  6. Ziche, M., Alessandri, G., and Gullino, P. M. (1989) Gangliosides promote the angiogenic response. Lab. Invest. 61, 629ā€“634.

    CASĀ  PubMedĀ  Google ScholarĀ 

  7. Ziche, M., Morbidelli, L., Masini, E., Amerini, S., Granger, H. J., Maggi, C. A., et al. (1994) Nitric oxide mediates angiogenesis in vivo and endothelial cell growth and migration in vitro promoted by substance P. J. Clin. Invest. 94, 2036ā€“2044.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  8. Ziche, M., Morbidelli, L., Choudhuri, R., Zhang, H. T., Donnini, S., Granger, H. J., and Bicknell, R. (1997) Nitric oxide-synthase lies downstream of vascular endothelial growth factor but not basic fibroblast growth factor induced angiogenesis. J. Clin. Invest. 99, 2625ā€“2634.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  9. Ziche, M., Maglione, D., Ribatti, D., Morbidelli, L., Lago, C. T., Battisti, M., et al. (1997) Placenta growth factor-1 (PlGF-1) is chemotactic, mitogenic and angiogenic. Lab. Invest. 76, 517ā€“531.

    CASĀ  PubMedĀ  Google ScholarĀ 

  10. Muthukkaruppan, V. and Auerbach, R. (1979) Angiogenesis in the mouse cornea. Science 206, 1416ā€“1418.

    ArticleĀ  Google ScholarĀ 

  11. Chen, C., Parangi, S., Tolentino, M. T., and Folkman, J. (1995) A strategy to discover circulating angiogenesis inhibitors generated by human tumors. Cancer Res. 55, 4230ā€“4233.

    CASĀ  PubMedĀ  Google ScholarĀ 

  12. Voest, E. E., Kenyon, B. M., Oā€™Reilly, M. S., Truitt, G., Dā€™Amato, R. J., and Folkman, J. (1995) Inhibition of angiogenesis in vivo by interleukin 12. J. Natl. Cancer Inst. 87, 581ā€“586.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  13. Polverini, P. J. and Leibovich, S. J. (1984) Induction of neovascularization in vivo and endothelial cell proliferation in vitro by tumor associated macrophages. Lab. Invest. 51, 635ā€“642.

    CASĀ  PubMedĀ  Google ScholarĀ 

  14. Ziche, M., Morbidelli, L., Pacini, M., Geppetti, P., Alessandri, G., and Maggi, C. A. (1990) Substance P stimulates neovascularization in vivo and proliferation of cultured endothelial cells. Microvasc. Res. 40, 264ā€“278.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  15. Bussolino, F., Ziche, M., Wang, J.M., Alessi, D., Morbidelli, L., Cremona, O., et al. (1991) In vitro and in vivo activation of endothelial cells by colony-stimulating factors. J. Clin. Invest. 87, 986ā€“995.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  16. Bussolino, F., Di Renzo, M. F., Ziche, M., Bocchietto, E., Oliviero, M., Naldini, L., et al. (1992) Hepatocyte growth factor is a potent angiogenic factor which stimulates endothelial cell motility and growth. J. Cell Biol. 119, 629ā€“641.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  17. Albini, A., Benelli, R., Presta, M., Rusnati, M., Ziche, M., Rubartelli, A., et al. (1996) HIV-tat protein is a heparin-binding angiogenic growth factor. Oncogene 12, 289ā€“297.

    CASĀ  PubMedĀ  Google ScholarĀ 

  18. Zhang, H. T., Kraft, P., Scott, P. A. E., Ziche, M., Weich, H. A., Harris, A. L., and Bicknell R. (1995) Enhancement of tumour growth and vascular density by transfection of vascular endothelial cell growth factor (VEGF) into MCF7 human breast carcinoma cells. J. Natl. Cancer Inst. 87, 213ā€“219.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  19. Coltrini, D., Gualandris, A., Nelli, E.M., Parolini, S., Molinari-Tosatti, M.P., Quarto, N., et al. (1995) Growth advantage and vascularization induced by basic fibroblast growth factor over-expression in endometrial HEC-1-B cells: an export-dependent mechanism of action. Cancer Res. 55, 4729ā€“4738.

    CASĀ  PubMedĀ  Google ScholarĀ 

  20. Gualandris, A., Rusnati, M., Belleri, M., Nelli, E. M., Bastaki, M., Molinari Tosatti, M. P., et al. (1996) Basic fibroblast growth factor over-expression in mouse endothelial cells: an autocrine model of angiogenesis and angioproliferative diseases. Cell Growth Differentiation 7, 147ā€“160.

    CASĀ  PubMedĀ  Google ScholarĀ 

  21. Ziche, M., Donnini, S., Morbidelli, L., Parenti, A., Gasparini, G., and Ledda, F. (1998) Linomide blocks angiogenesis by breast carcinoma vascular endothelial growth factor transfectants. Br. J. Cancer 77, 1123ā€“1129.

    ArticleĀ  CASĀ  PubMedĀ  Google ScholarĀ 

  22. Choudhuri, R., Zhang, H. T., Donnini, S., Ziche, M., and Bicknell, R. (1997) An angiogenic role for the neurotrophins midkine and pleiotrophin in tumorigenesis. Cancer Res. 57, 1814ā€“1819.

    CASĀ  PubMedĀ  Google ScholarĀ 

  23. Proia, A. D., Chadler, D. B., Haynes, W. L., Smith, C. F., Suvarenamani, C., Erkel, F. H., and Klintworth, G. K. (1988) Quantification of corneal neovascularization using computerized image analysis. Lab. Invest. 58, 473-479.

    Google ScholarĀ 

  24. Conrad, T. J., Chandler, D. B., Corless, J. M., and Klintworth, G. K. (1994) In vivo measurement of corneal angiogenesis with video data acquisition and computerized image analysis. Lab. Invest. 70, 426ā€“434.

    CASĀ  PubMedĀ  Google ScholarĀ 

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Ā© 2001 Humana Press Inc.

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Ziche, M. (2001). Corneal Assay for Angiogenesis. In: Murray, J.C. (eds) Angiogenesis Protocols. Methods in Molecular Medicineā„¢, vol 46. Humana Press. https://doi.org/10.1385/1-59259-143-4:131

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  • DOI: https://doi.org/10.1385/1-59259-143-4:131

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-698-7

  • Online ISBN: 978-1-59259-143-5

  • eBook Packages: Springer Protocols

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