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Standardization of the HET-CAM Assay (Hen´s Egg Test – Chorionallantoic Membrane) for Validation of New Biomedical Applications.

Project Aim

The goal of this project is to investigate the impact of angiogenesis in neoplastic diseases and other angiogenic disorders. A test system for this objective has to be developed, which allows quantification of angiogenesis and is subsequently capable of providing adequate complimentary animal models so that a large number of these animal experiments can be abandoned.

Project Background

As a test organ, the membrane located directly underneath the shell of an incubated chicken egg (chorionallantois membrane) is used. It contains veins and arteries but no nerves. A large part of the experiments can be completed prior to the closing of the neural tube on the 11th day of incubation, so that we can assume that only minimal stress is exhibited on the embryo. The HET-CAM assay is consequently not classified as an animal experiment. The test substance is applied onto the chorionallantois membrane and the subsequent reaction (morphological changes in vessels, hemorrhaging, vaso activity, etc.) is monitored. By modification of the HET-CAM-assay, it is possible to test various materials for their biocompatibility which will eventually be used for medical implants or prosthesis. The HET-CAM assay can even be used in a modified version (1) for applications in the area of cell transplantation experiments or tissue engineering. The chorionallantois membrane is also suited for pharmacological tests of vaso activity as well as thrombogenic and thrombolytic effects. Investigations on the metabolism of test substances are furthermore possible.
Tumor-angiogenesis is currently being investigated with the aid of a modified HET-CAM assay. Most tumors induce vessel formation via messengers that leads to the immigration of the vessels into the tumor which provides it with nutrients – a process which is called angiogenesis. Angiogenesis is a very complex process, which is not (yet) fully understood in detail, but it contains, among other things, the disaggregation of the basal lamina, to provide the immigration of new endothelial cells into the tumor via existing vessels. Anti-angiogenic therapies which inhibit angiogenesis and consequently tumor growth are currently seen as promising additions to chemotherapy in the treatment of cancer.

Experiment Status

The zet-LSL currently investigates the growth properties of tumor cells onto the CAM in cooperation with selected institutes of biomedical research at the Medical University of Vienna and the biotechnology company, Baxter AG. Currently, different materials are additionally being tested with the HET-CAM assay for their biocompatibility. They could possibly be utilized in the future for setting drainage devices in the human eye for Glaucoma treatment.

Start of the Study

1 June 2006

Participating Laboratories

Medical University of Vienna/ Vienna General Hospital – specific institutes of biomedical research, zet-LSL.

Financing

Pollux Foundation

References

(1) Eder C, Falkner E, Nehrer S, Losert UM, and Schoeffl H: Introducing the concept of the 3Rs into tissue engineering research. ALTEX. 23: 17-23, 2006.