Spontaneous metastasis assay

Box GM, Eccles SA (2011) Simple experimental and spontaneous metastasis assays in mice. Methods Mol Biol 769 311-329... 

Spontaneous Metastasis Assay from a Primary Mammary Fat Pad Tumor Resection... 

There are so many variations of the spontaneous metastasis assay that it is impossible to describe all of them here. Therefore, two approaches are provided as examples from which the reader is to... 

The mechanics for experimental metastasis assays are similar to those for the spontaneous metastasis assay. As above, volume and viscosity of the inoculum are important parameters. Volumes should never exceed 0.2 ml because plasma blood volume exceeds the normal range ( 10%) such that the distribution pattern of cells is altered. Tumor density is even more critical for intravascular injections since introduction of emboli can cause death because of vascular obstruction. Therefore, the same concentration maximums reported above are also applicable for this type of assay. 

Experimental pulmonary metastasis was assessed by tumor cell injection into the lateral tail vein of mice. C57BL/6 mice were injected intravenously (IV) with B16-BL6 melanoma (5 X 10 ) admixed with or without chitin derivatives. The mice were killed 14 days after tumor inoculation, and the number of lung tumor colonies was counted. In a spontaneous pulmonary metastasis assay, mice were inoculated subcutaneously with B16-BL6 cells (5 X 10 ) in the right hind footpad. The primary tumors were surgically removed on day 21 after tumor inoculation. The injection of chitin derivatives was performed on various days before or after the amputation. Mice were killed 14 days after the amputation, and the number of lung tumor colonies was counted. An outline of the experiment is shown in Scheme 3. 

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