title: The in ovo CAM-assay as a xenograft model for sarcoma.
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.
authors.affiliation: Department of Orthopaedic Surgery and Traumatology, Ghent University Hospital. Gwen.Sys@Ugent.be
authors.forename: Gwen M L
authors.initials: GM
authors.lastname: Sys
authors.forename: Lore
authors.initials: L
authors.lastname: Lapeire
authors.forename: Nikita
authors.initials: N
authors.lastname: Stevens
authors.forename: Herman
authors.initials: H
authors.lastname: Favoreel
authors.forename: Ramses
authors.initials: R
authors.lastname: Forsyth
authors.forename: Marc
authors.initials: M
authors.lastname: Bracke
authors.forename: Olivier
authors.initials: O
authors.lastname: De Wever
mesh_terms: D000818:Animals; D001859:Bone Neoplasms / Q000473:pathology / Q000601:surgery; D045744:Cell Line, Tumor; D002642:Chick Embryo; D002813:Chondrosarcoma / Q000473:pathology / Q000601:surgery; D049033:Chorioallantoic Membrane / Q000473:pathology* / Q000601:surgery*; D006801:Humans; D009368:Neoplasm Transplantation / Q000379:methods*; D012516:Osteosarcoma / Q000473:pathology / Q000601:surgery; D012509:Sarcoma / Q000473:pathology* / Q000601:surgery; D014183:Transplantation, Heterologous / Q000379:methods*; D023041:Xenograft Model Antitumor Assays
references.citation: Nat Med. 2011 Oct 23;17(11):1514-20
references.pmid: 22019887
references.citation: Proc Natl Acad Sci U S A. 2005 Feb 1;102(5):1643-8
references.pmid: 15665100
references.citation: Cell. 2011 Mar 4;144(5):646-74
references.pmid: 21376230
references.citation: Int J Mol Sci. 2012;13(8):9959-70
references.pmid: 22949841
references.citation: Int J Dev Biol. 2010;54(5):887-96
references.pmid: 19757378
references.citation: J Vis Exp. 2009 Nov 30;(33):
references.pmid: 19949373
references.citation: Cancer Res. 1982 May;42(5):1826-37
references.pmid: 7066899
references.citation: Clin Cancer Res. 1999 Aug;5(8):2198-204
references.pmid: 10473106
references.citation: Anticancer Drugs. 2011 Oct;22(9):827-41
references.pmid: 21623183
references.citation: Histochem Cell Biol. 2008 Dec;130(6):1119-30
references.pmid: 19005674
references.citation: J Natl Cancer Inst. 2010 Jun 16;102(12):866-80
references.pmid: 20484105
references.citation: Br J Cancer. 1977 Mar;35(3):347-56
references.pmid: 856240
references.citation: J Clin Oncol. 2010 Apr 1;28(10):1796-805
references.pmid: 20194847
references.citation: Am J Pathol. 1975 Jun;79(3):597-618
references.pmid: 1094838
references.citation: Adv Drug Deliv Rev. 2007 Sep 30;59(11):1162-76
references.pmid: 17870202
references.citation: Lab Invest. 2003 Jun;83(6):877-87
references.pmid: 12808123
references.citation: Clin Orthop Relat Res. 2004 Dec;(429):286-91
references.pmid: 15577500
references.citation: Cancer Discov. 2011 Nov;1(6):508-23
references.pmid: 22586653
references.citation: CA Cancer J Clin. 2001 Jan-Feb;51(1):15-36
references.pmid: 11577478
references.citation: BMC Res Notes. 2010 Mar 04;3:58
references.pmid: 20202196
references.citation: Cancer Lett. 2012 Dec 29;326(1):69-78
references.pmid: 22841668
references.citation: Nat Rev Cancer. 2011 Jul 14;11(8):541-57
references.pmid: 21753790
references.citation: Cancer Cell. 2012 Mar 20;21(3):309-22
references.pmid: 22439926
references.citation: Proc Natl Acad Sci U S A. 2010 May 4;107(18):8352-6
references.pmid: 20404174
references.citation: Nat Protoc. 2007;2(3):504-11
references.pmid: 17406614
references.citation: Mayo Clin Proc. 2007 Nov;82(11):1409-32
references.pmid: 17976362
references.citation: Nat Rev Clin Oncol. 2012 Apr 17;9(6):338-50
references.pmid: 22508028