Intravenous Injection of Cancer Cells into Chicken Chorioallantoic Membrane Vasculature: A Procedure to Establish Chorioallantoic Membrane Cancer Model to Study Cancer Invasion and Metastasis
Trascrizione
The chicken chorioallantoic membrane, or CAM, of a fertilized chicken egg is the extra-embryonic membrane supported by a rich vascular network. This highly vascularized membrane lining the non-vascular shell membrane facilitates the exchange of gases and nutrients with the developing embryo.
To generate a CAM cancer model in vitro, begin with a viable, fertilized, shell-less chicken embryo bearing a fully developed CAM in a dish.
Next, assemble the injection apparatus containing a tubing attached to a syringe carrying a single-cell suspension of transduced cancer cells in a suitable chilled buffer. Connect a customized glass needle to the tubing for high precision delivery during subsequent injection.
Now, transfer the embryo under a microscope. Identify a suitable vein close to the bifurcation point on the CAM surface. Inject the cancer cell suspension into the selected vein and ensure complete delivery. Gently retract the needle. Cover the embryo and incubate it under appropriate conditions.
The dense vasculature within the mesodermal layer harbors the injected tumor cells, facilitating primary tumor formation. Eventually, some of these cells migrate through the vasculature and extravasate, invading the surrounding stroma, where they proliferate to form metastatic foci.
