Skip to main content
SpringerLink
Log in

Methods for Analyzing Tumor Angiogenesis in the Chick Chorioallantoic Membrane Model

  • Protocol
  • First Online:
Breast Cancer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1406))

Abstract

Models of tumor angiogenesis have played a critical role in understanding the mechanisms involved in the recruitment of vasculature to the tumor mass, and have also provided a platform for testing antiangiogenic potential of new therapeutics that combat the development of malignant growth. In this regard, the chorioallantoic membrane (CAM) of the developing chick embryo has proven to be an elegant model for investigation of angiogenic processes. Here, we describe methods for effectively utilizing the preestablished vascular network of the chick CAM to investigate and quantify tumor-associated angiogenesis in a breast tumor model.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Bader AG, Kang S, Vogt PK (2006) Cancer-specific mutations in PIK3CA are oncogenic in vivo. Proc Natl Acad Sci U S A 103(5):1475–1479

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  2. Eliceiri BP, Klemke R, Stromblad S, Cheresh DA (1998) Integrin avb3 requirement for sustained mitogen-activated protein kinase activity during angiogenesis. J Cell Biol 140(5):1255–1263

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  3. Hanahan D, Folkman J (1996) Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell 86:353–364

    Article  PubMed  CAS  Google Scholar 

  4. Liu M, Scanlon CS, Banerjee R, Russo N, Inglehart RC, Willis AL, Weiss SJ, D’Silva NJ (2013) The histone methyltransferase EZH2 mediates tumor progression on the chick chorioallantoic membrane assay, a novel model of head and neck squamous cell carcinoma. Transl Oncol 6(3):273–281

    Article  PubMed  PubMed Central  Google Scholar 

  5. Zijlstra A, Seandel M, Kupriyanova TA, Partridge JJ, Madsen MA, Hahn-Dantona EA, Quigley JP, Deryugina EI (2006) Proangiogenic role of neutrophil-like inflammatory heterophils during neovascularization induced by growth factors and human tumor cells. Blood 107(1):317–327

    Article  PubMed  PubMed Central  Google Scholar 

  6. Debefve E, Pegaz B, Ballini JP, van den Bergh H (2009) Combination therapy using verteporfin and ranibizumab; optimizing the timing in the CAM model. Photochem Photobiol 85(6):1400–1408

    Article  PubMed  CAS  Google Scholar 

  7. Palacios-Arreola MI, Nava-Castro KE, Castro JI, Garcia-Zepeda E, Carrero JC, Morales-Montor J (2014) The role of chemokines in breast cancer pathology and its possible use as therapeutic targets. J Immunol Res 2014:849720

    Article  PubMed  PubMed Central  Google Scholar 

  8. Ucuzian AA, Greisler HP (2007) In vitro models of angiogenesis. World J Surg 31(4):654–663

    Article  PubMed  Google Scholar 

  9. Bai Y, Zhao M, Zhang C, Li S, Qi Y, Wang B, Huang L, Li X (2014) Anti-angiogenic effects of a mutant endostatin: a new prospect for treating retinal and choroidal neovascularization. PLoS One 9(11):e112448

    Article  PubMed  PubMed Central  Google Scholar 

  10. Nicosia RF (2009) The aortic ring model of angiogenesis: a quarter century of search and discovery. J Cell Mol Med 13(10):4113–4136

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  11. Couffinhal T, Dufourcq P, Barandon L, Leroux L, Duplaa C (2009) Mouse models to study angiogenesis in the context of cardiovascular diseases. Front Biosci 14:3310–3325

    Article  CAS  Google Scholar 

  12. Norrby K (2006) In vivo models of angiogenesis. J Cell Mol Med 10(3):588–612

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  13. Tredan O, Lacroix-Triki M, Guiu S, Mouret-Reynier MA, Barriere J, Bidard FC, Braccini AL, Mir O, Villanueva C, Barthelemy P (2015) Angiogenesis and tumor microenvironment: bevacizumab in the breast cancer model. Target Oncol 10:189

    Article  PubMed  Google Scholar 

  14. Mittal K, Ebos J, Rini B (2014) Angiogenesis and the tumor microenvironment: vascular endothelial growth factor and beyond. Semin Oncol 41(2):235–251

    Article  PubMed  CAS  Google Scholar 

  15. Armstrong PB, Quigley JP, Sidebottom E (1982) Transepithelial invasion and intramesenchymal infiltration of the chick embryo chorioallantois by tumor cell lines. Cancer Res 42(5):1826–1837

    PubMed  CAS  Google Scholar 

  16. Kain KH, Miller JW, Jones-Paris CR, Thomason RT, Lewis JD, Bader DM, Barnett JV, Zijlstra A (2014) The chick embryo as an expanding experimental model for cancer and cardiovascular research. Dev Dyn 243(2):216–228

    Article  PubMed  PubMed Central  Google Scholar 

  17. Nguyen M, Shing Y, Folkman J (1994) Quantitation of angiogenesis and antiangiogenesis in the chick embryo chorioallantoic membrane. Microvasc Res 47(1):31–40

    Article  PubMed  CAS  Google Scholar 

  18. Nowak-Sliwinska P, Segura T, Iruela-Arispe ML (2014) The chicken chorioallantoic membrane model in biology, medicine and bioengineering. Angiogenesis 17(4):779–804

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  19. Ribatti D (2012) Chicken chorioallantoic membrane angiogenesis model. Methods Mol Biol 843:47–57

    Article  PubMed  CAS  Google Scholar 

  20. Ribatti D (2014) The chick embryo chorioallantoic membrane as a model for tumor biology. Exp Cell Res 328:314

    Article  PubMed  CAS  Google Scholar 

  21. Ausprunk DH, Knighton DR, Folkman J (1974) Differentiation of vascular endothelium in the chick chorioallantois: a structural and autoradiographic study. Dev Biol 38:237–247

    Article  PubMed  CAS  Google Scholar 

  22. Borges J, Tegtmeier FT, Pardon NT, Mueller MC, Lang EM, Stark B (2003) Chorioallantoic membrane angiogenesis model for tissue engineering: a new twist on a classic model. Tissue Eng 19(3):441–450

    Article  Google Scholar 

  23. Brooks PC, Montgomery AM, Cheresh DA (1999) Use of the 10-day-old chick embryo model for studying angiogenesis. Methods Mol Biol 129:257–269

    PubMed  CAS  Google Scholar 

  24. Ribatti D, Nico B, Vacca A, Roncali L, Burri PH, Djonov V (2001) Chorioallantoic membrane capillary bed: a useful target for studying angiogenesis and anti-angiogenesis in vivo. Anat Rec 264(4):317–324

    Article  PubMed  CAS  Google Scholar 

  25. Tay SL, Heng PW, Chan LW (2012) The CAM-LDPI method: a novel platform for the assessment of drug absorption. J Pharm Pharmacol 64(4):517–529

    Article  PubMed  CAS  Google Scholar 

  26. Tay SL, Heng PW, Chan LW (2012) The chick chorioallantoic membrane imaging method as a platform to evaluate vasoactivity and assess irritancy of compounds. J Pharm Pharmacol 64(8):1128–1137

    Article  PubMed  CAS  Google Scholar 

  27. Janković BD, Isaković K, Lukić ML, Vujanović NL, Petrović S, Marković BM (1975) Immunological capacity of the chicken embryo. Immunology 29(3):497–508

    PubMed  PubMed Central  Google Scholar 

  28. Murphy JB (1914) Factors of resistance to heteroplastic tissue-grafting: studies in tissue specificity III. J Exp Med 19:513–522

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  29. Murphy JB (1914) The ultimate fate of mammalian tissue implanted in the chick embryo II. J Exp Med 19:181–186

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  30. Ausprunk D, Knighton DR, Folkman J (1975) Vascularization of normal and neoplastic tissues grafted to the chick chorioallantois. Am J Pathol 79:597–618

    PubMed  CAS  PubMed Central  Google Scholar 

  31. Dagg CP, Karnofsky DA, Roddy J (1956) Growth of transplantable human tumors in the chick embryo and hatched chick. Cancer Res 16(7):589–594

    PubMed  CAS  Google Scholar 

  32. Deryugina EI, Quigley JP (2008) Chick embryo chorioallantoic membrane model systems to study and visualize human tumor cell metastasis. Histochem Cell Biol 130(6):1119–1130

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  33. Easty GC, Easty DM, Tchao R (1969) The growth of heterologous tumour cells in chick embryos. Eur J Cancer 5:287–295

    Article  PubMed  CAS  Google Scholar 

  34. Fein MR, Egeblad M (2013) Caught in the act: revealing the metastatic process by live imaging. Dis Model Mech 6(3):580–593

    Article  PubMed  PubMed Central  Google Scholar 

  35. Fergelot P, Bernhard JC, Soulet F, Kilarski WW, Leon C, Courtois N, Deminiere C, Herber JM, Antczak P, Falciani F, Rioux-Leclercq N, Patard JJ, Ferriere JM, Ravaud A, Hagedorn M, Bikfalvi A (2013) The experimental renal cell carcinoma model in the chick embryo. Angiogenesis 16(1):181–194

    Article  PubMed  Google Scholar 

  36. Palmer TD, Lewis J, Zijlstra A (2011) Quantitative analysis of cancer metastasis using an avian embryo model. J Vis Exp (51): pii: 2815

    Google Scholar 

  37. van Beijnum JR, Nowak-Sliwinska P, van den Boezem E, Hautvast P, Buurman WA, Griffioen AW (2013) Tumor angiogenesis is enforced by autocrine regulation of high-mobility group box 1. Oncogene 32(3):363–374

    Article  PubMed  Google Scholar 

  38. Zijlstra A, Lewis J, Degryse B, Stuhlmann H, Quigley JP (2008) The inhibition of tumor cell intravasation and subsequent metastasis via regulation of in vivo tumor cell motility by the tetraspanin CD151. Cancer Cell 13(3):221–234

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  39. Isachenko V, Isachenko E, Mallmann P, Rahimi G (2013) Increasing follicular and stromal cell proliferation in cryopreserved human ovarian tissue after long-term precooling prior to freezing: in vitro versus chorioallantoic membrane (CAM) xenotransplantation. Cell Transplant 22(11):2053–2061

    Article  PubMed  Google Scholar 

  40. Lokman NA, Elder AS, Ricciardelli C, Oehler MK (2012) Chick chorioallantoic membrane (CAM) assay as an in vivo model to study the effect of newly identified molecules on ovarian cancer invasion and metastasis. Int J Mol Sci 13(8):9959–9970

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  41. Lopez-Rivera E, Jayaraman P, Parikh F, Davies MA, Ekmekcioglu S, Izadmehr S, Milton DR, Chipuk JE, Grimm EA, Estrada Y, Aquirre-Ghiso J, Sikora AG (2014) Inducible nitric oxide synthase drives mTOR pathway activation and proliferation of human melanoma by reversible nitrosylation of TSC2. Cancer Res 74(4):1067–1078

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  42. Mu X, Sultankulov B, Agarwal R, Mahjoub A, Schott T, Greco N, Huard J, Weiss K (2014) Chick embryo extract demethylates tumor suppressor genes in osteosarcoma cells. Clin Orthop Relat Res 472(3):865–873

    Article  PubMed  PubMed Central  Google Scholar 

  43. Contois LW, Akalu A, Caron JM, Tweedie E, Cretu A, Henderson T, Liaw L, Friese IR, Vary C, Brooks PC (2015) Inhibition of tumor-associated αvβ3 integrin regulates the angiogenic switch by enhancing expression of IGFBP-4 leading to reduced melanoma growth and angiogenesis in vivo. Angiogenesis 18:31

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  44. Downey CM, Singla AK, Villemaire ML, Buie HR, Boyd SK, Jirik FR (2012) Quantitative ex-vivo micro-computed tomographic imaging of blood vessels and necrotic regions within tumors. PLoS One 7(7):e41685

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  45. Mondy WL, Cameron D, Timmermans JP, De Clerck N, Sasov AC, Casteleyn C, Piegl LA (2009) Micro-CT or corrosion casts for use in the computer-aided design of microvasculature. Tissue Eng Part C Methods 15:729–738

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported in part by a grant from the Maine Cancer Foundation (MCF) to PCB, NIH Center for Biomedical Research Excellence P30GM103392 (P.I. Robert Friesel), and institutional support from the Maine Medical Center.

Author information

Authors and Affiliations

  1. Center for Molecular Medicine, Maine Medical Center Research Institute, 81 Research Drive, Scarborough, ME, 04074, USA

    Jacquelyn J. Ames, Terry Henderson, Lucy Liaw & Peter C. Brooks Ph.D.

  2. Graduate School of Biomedical Science and Engineering, University of Maine, Orono, ME, 04469, USA

    Jacquelyn J. Ames

Authors
  1. Jacquelyn J. Ames
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Terry Henderson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lucy Liaw
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Peter C. Brooks Ph.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Peter C. Brooks Ph.D. .

Editor information

Editors and Affiliations

  1. School of Medicine, Stony Brook University, Stony Brook, New York, USA

    Jian Cao

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer Science+Business Media New York

About this protocol

Cite this protocol

Ames, J.J., Henderson, T., Liaw, L., Brooks, P.C. (2016). Methods for Analyzing Tumor Angiogenesis in the Chick Chorioallantoic Membrane Model. In: Cao, J. (eds) Breast Cancer. Methods in Molecular Biology, vol 1406. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3444-7_22

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-3444-7_22

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3442-3

  • Online ISBN: 978-1-4939-3444-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation