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Updated: May 9, 2026
The In ovo CAM-assay as a Xenograft Model for Sarcoma
Published on: July 17, 2013
The in ovo CAM-assay as a xenograft model for sarcoma
Gwen M L Sys1, Lore Lapeire, Nikita Stevens
1Department of Orthopaedic Surgery and Traumatology, Ghent University Hospital. Gwen.Sys@Ugent.be
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.
Insights
A novel chick chorioallantoic membrane xenotransplant model offers a low-cost, reproducible method for studying sarcoma. This model aids in personalized medicine by enabling detailed analysis of tumor growth and host interactions.
Area of Science:
- Oncology
- Developmental Biology
- Tissue Engineering
Background:
- Sarcoma is a rare and heterogeneous cancer, hindering therapeutic development.
- Personalized medicine requires effective patient stratification, necessitating robust preclinical models.
- Existing models often lack reproducibility and affordability for sarcoma research.
Purpose of the Study:
- To establish and validate a reproducible, low-cost chick chorioallantoic membrane (CAM) xenotransplant model for sarcoma.
- To assess the utility of the CAM model for studying sarcoma biology and host-tumor interactions.
- To provide a detailed protocol for in ovo grafting of sarcoma tissues and cell lines.
Main Methods:
- In ovo grafting of fresh sarcoma tumor tissues and single-cell suspensions onto the chick chorioallantoic membrane.
- Utilizing fluorescently labeled sarcoma cell lines (Saos-2, SW1353) for in vivo tracking.
- Employing ECM gel for localized grafting of single-cell suspensions.
- Assessing graft viability, proliferation (Ki67), necrosis, infiltration, and host responses (fibroblast infiltration, vascular ingrowth) via microscopy and histology.
Main Results:
- Achieved chick survival rates up to 75% with the CAM xenotransplant model.
- Demonstrated the model's capability to analyze graft behavior, including viability, proliferation, necrosis, and infiltration.
- Observed host responses such as fibroblast infiltration and vascular ingrowth.
- Found that Ki67 proliferation index correlates with distance from the CAM surface and duration of engraftment.
Conclusions:
- The chick chorioallantoic membrane provides a viable, immunodeficient host for sarcoma xenotransplantation.
- This model is suitable for studying sarcoma biology, host-tumor interactions, and evaluating therapeutic strategies.
- The protocol facilitates reproducible, low-cost research crucial for advancing personalized medicine in sarcoma.
