The In ovo CAM-assay as a Xenograft Model for Sarcoma |
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Authors: | Gwen M.L. Sys Lore Lapeire Nikita Stevens Herman Favoreel Ramses Forsyth Marc Bracke Olivier De Wever |
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Affiliation: | 1.Department of Orthopaedic Surgery and Traumatology, Ghent University Hospital;2.Department of Radiation Oncology and Experimental Cancer Research, Ghent University;3.Department of Virology, Parasitology, and Immunology, Ghent University;4.Pathlicon |
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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. |
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Keywords: | Cancer Biology Issue 77 Medicine Cellular Biology Molecular Biology Biomedical Engineering Bioengineering Developmental Biology Anatomy Physiology Oncology Surgery Adipose Tissue Connective Tissue Neoplasm Muscle Tissue Sarcoma Animal Experimentation Cell Culture Techniques Neoplasms Experimental Neoplasm Transplantation Biological Assay Sarcomas CAM-assay CAM assay xenograft proliferation invasion cancer tumor in ovo animal model |
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