The in Ovo CAM-assay As a Xenograft Model for Sarcoma

Overview

Journal J Vis Exp

Specialties Biology
General Medicine

Date 2013 Jul 30

PMID 23892612

Citations 36

Authors

Gwen M L Sys

Lore Lapeire

Nikita Stevens

Herman Favoreel

Ramses Forsyth

Marc Bracke

Olivier de Wever

Affiliations

Soon will be listed here.

Abstract

Sarcoma is a very rare disease that is heterogeneous in nature, all hampering the development of new therapies. Sarcoma patients are ideal candidates for personalized medicine after stratification, explaining the current interest in developing a reproducible and low-cost xenotransplant model for this disease. The chick chorioallantoic membrane is a natural immunodeficient host capable of sustaining grafted tissues and cells without species-specific restrictions. In addition, it is easily accessed, manipulated and imaged using optical and fluorescence stereomicroscopy. Histology further allows detailed analysis of heterotypic cellular interactions. This protocol describes in detail the in ovo grafting of the chorioallantoic membrane with fresh sarcoma-derived tumor tissues, their single cell suspensions, and permanent and transient fluorescently labeled established sarcoma cell lines (Saos-2 and SW1353). The chick survival rates are up to 75%. The model is used to study graft- (viability, Ki67 proliferation index, necrosis, infiltration) and host (fibroblast infiltration, vascular ingrowth) behavior. For localized grafting of single cell suspensions, ECM gel provides significant advantages over inert containment materials. The Ki67 proliferation index is related to the distance of the cells from the surface of the CAM and the duration of application on the CAM, the latter determining a time frame for the addition of therapeutic products.

Citing Articles

Impact of late-stage hypoxic stimulation and layer breeder age on embryonic development, hatching and chick quality.

Agbehadzi R, Kumi G, Adjei-Mensah B, Hamidu J, Tona K Poult Sci. 2024; 104(2):104691.

PMID: 39731869 PMC: 11748728. DOI: 10.1016/j.psj.2024.104691.

Xenografting Human Musculoskeletal Sarcomas in Mice, Chick Embryo, and Zebrafish: How to Boost Translational Research.

Giusti V, Miserocchi G, Sbanchi G, Pannella M, Hattinger C, Cesari M Biomedicines. 2024; 12(8).

PMID: 39200384 PMC: 11352184. DOI: 10.3390/biomedicines12081921.

Enhancing maturity in 3D kidney micro-tissues through clonogenic cell combinations and endothelial integration.

Abdollahzadeh F, Khoshdel-Rad N, Bahrehbar K, Erfanian S, Ezzatizadeh V, Totonchi M J Cell Mol Med. 2024; 28(11):e18453.

PMID: 38818569 PMC: 11140233. DOI: 10.1111/jcmm.18453.

Experimental Tumor Induction and Evaluation of Its Treatment in the Chicken Embryo Chorioallantoic Membrane Model: A Systematic Review.

Mesas C, Chico M, Doello K, Lara P, Moreno J, Melguizo C Int J Mol Sci. 2024; 25(2).

PMID: 38255911 PMC: 10815318. DOI: 10.3390/ijms25020837.

Technical Implications of the Chicken Embryo Chorioallantoic Membrane Assay to Elucidate Neuroblastoma Biology.

Patino-Morales C, Jaime-Cruz R, Ramirez-Fuentes T, Villavicencio-Guzman L, Salazar-Garcia M Int J Mol Sci. 2023; 24(19).

PMID: 37834193 PMC: 10572838. DOI: 10.3390/ijms241914744.

References

Derose Y, Wang G, Lin Y, Bernard P, Buys S, Ebbert M . Tumor grafts derived from women with breast cancer authentically reflect tumor pathology, growth, metastasis and disease outcomes. Nat Med. 2011; 17(11):1514-20. PMC: 3553601. DOI: 10.1038/nm.2454. View

Hagedorn M, Javerzat S, Gilges D, Meyre A, de Lafarge B, Eichmann A . Accessing key steps of human tumor progression in vivo by using an avian embryo model. Proc Natl Acad Sci U S A. 2005; 102(5):1643-8. PMC: 547849. DOI: 10.1073/pnas.0408622102. View

Hanahan D, Weinberg R . Hallmarks of cancer: the next generation. Cell. 2011; 144(5):646-74. DOI: 10.1016/j.cell.2011.02.013. View

Lokman N, Elder A, Ricciardelli C, Oehler M . 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. 2012; 13(8):9959-9970. PMC: 3431839. DOI: 10.3390/ijms13089959. View

de Wever O, Hendrix A, De Boeck A, Westbroek W, Braems G, Emami S . Modeling and quantification of cancer cell invasion through collagen type I matrices. Int J Dev Biol. 2009; 54(5):887-96. DOI: 10.1387/ijdb.092948ow. View

Dohle D, Pasa S, Gustmann S, Laub M, Wissler J, Jennissen H . Chick ex ovo culture and ex ovo CAM assay: how it really works. J Vis Exp. 2009; (33). PMC: 3157849. DOI: 10.3791/1620. View

Armstrong P, Quigley J, Sidebottom E . Transepithelial invasion and intramesenchymal infiltration of the chick embryo chorioallantois by tumor cell lines. Cancer Res. 1982; 42(5):1826-37. View

Hoffmann J, SCHMIDT-PETER P, Hansch W, Naundorf H, Bunge A, Becker M . Anticancer drug sensitivity and expression of multidrug resistance markers in early passage human sarcomas. Clin Cancer Res. 1999; 5(8):2198-204. View

Decaudin D . Primary human tumor xenografted models ('tumorgrafts') for good management of patients with cancer. Anticancer Drugs. 2011; 22(9):827-41. DOI: 10.1097/CAD.0b013e3283475f70. View

10.

Deryugina E, Quigley J . Chick embryo chorioallantoic membrane model systems to study and visualize human tumor cell metastasis. Histochem Cell Biol. 2008; 130(6):1119-30. PMC: 2699943. DOI: 10.1007/s00418-008-0536-2. View

11.

Hendrix A, Maynard D, Pauwels P, Braems G, Denys H, Van den Broecke R . Effect of the secretory small GTPase Rab27B on breast cancer growth, invasion, and metastasis. J Natl Cancer Inst. 2010; 102(12):866-80. PMC: 2886092. DOI: 10.1093/jnci/djq153. View

12.

Knighton D, Ausprunk D, Tapper D, Folkman J . Avascular and vascular phases of tumour growth in the chick embryo. Br J Cancer. 1977; 35(3):347-56. PMC: 2025276. DOI: 10.1038/bjc.1977.49. View

13.

Nielsen T, West R . Translating gene expression into clinical care: sarcomas as a paradigm. J Clin Oncol. 2010; 28(10):1796-805. DOI: 10.1200/JCO.2009.26.1917. View

14.

Ausprunk D, Knighton D, Folkman J . Vascularization of normal and neoplastic tissues grafted to the chick chorioallantois. Role of host and preexisting graft blood vessels. Am J Pathol. 1975; 79(3):597-618. PMC: 1913018. View

15.

Vargas A, Zeisser-Labouebe M, Lange N, Gurny R, Delie F . The chick embryo and its chorioallantoic membrane (CAM) for the in vivo evaluation of drug delivery systems. Adv Drug Deliv Rev. 2007; 59(11):1162-76. DOI: 10.1016/j.addr.2007.04.019. View

16.

Gil-Benso R, Lopez-Gines C, Lopez-Guerrero J, Carda C, Callaghan R, Navarro S . Establishment and characterization of a continuous human chondrosarcoma cell line, ch-2879: comparative histologic and genetic studies with its tumor of origin. Lab Invest. 2003; 83(6):877-87. DOI: 10.1097/01.lab.0000073131.34648.ea. View

17.

Mankin H, Hornicek F, Rosenberg A, Harmon D, Gebhardt M . Survival data for 648 patients with osteosarcoma treated at one institution. Clin Orthop Relat Res. 2004; (429):286-91. DOI: 10.1097/01.blo.0000145991.65770.e6. View

18.

Bertotti A, Migliardi G, Galimi F, Sassi F, Torti D, Isella C . A molecularly annotated platform of patient-derived xenografts ("xenopatients") identifies HER2 as an effective therapeutic target in cetuximab-resistant colorectal cancer. Cancer Discov. 2012; 1(6):508-23. DOI: 10.1158/2159-8290.CD-11-0109. View

19.

Greenlee R, Murray T, Thun M . Cancer statistics, 2001. CA Cancer J Clin. 2001; 51(1):15-36. DOI: 10.3322/canjclin.51.1.15. View

20.

Balke M, Neumann A, Kersting C, Agelopoulos K, Gebert C, Gosheger G . Morphologic characterization of osteosarcoma growth on the chick chorioallantoic membrane. BMC Res Notes. 2010; 3:58. PMC: 2838906. DOI: 10.1186/1756-0500-3-58. View