Use of NeoR B16F1 murine melanoma cells to assess clonality of experimental metastases in the immune-deficient chick embryo

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Abstract
Recent work on molecular and genetic aspects of metastasis has emphasized the need for assays in immune-deficient animal hosts. The commonly used assays in athymic nude mice may not always be appropriate, and assays in other hosts are required. We have developed a metastasis assay in the naturally immune-deficient chicken embryo. As part of our characterization of this assay we have examined the clonality of individual experimental (i.v.-derived) metastases in this host. For these studies we developed a cell line, B16-Neo, from parental B16F1 murine melanoma cells. B16-Neo cells carry a stable drug-resistance marker, the bacterialneo gene, which confers resistance to the drug G418, but are unaltered in experimental metastatic properties in the chick embryo relative to parental B16F1 cells. We observe that the majority of individual liver tumors that arise following i.v. injection of mixtures of these cells contain cells of a single marker phenotype and are likely to be clonal in origin. These results are similar to those obtained by others for metastases in immune-competent mice, suggesting similar mechanisms of metastasis formation in these two systems. In both hosts it should be noted, however, that a small but significant proportion of metastases appearnot to be clonal in origin.