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Characteristics of Cultured Human Melanocytes Isolated from Different Stages of Tumor Progression¹

The Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104 [M. H., J. T., J. L. B., D. H., H. K.], and the Pigmented Lesion Study Group [D. E. E., E. B., D. G., W. H. C.] and the Departments of Human Genetics [G. B.] and Pathology and Laboratory Medicine [P. N.], University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104

Normal melanocytes and melanocytes of normal nevi, primary melanoma in the radial (RGP) and vertical (VGP) growth phases, and metastatic melanoma exhibited and maintained phenotypic differences when grown in tissue culture or in experimental animals. Only metastatic and VGP primary melanoma cells were tumorigenic in athymic nude mice and had nonrandom chromosomal abnormalities involving chromosomes 1, 6, and 7. The colony-forming efficiency in soft agar was also highest in these two cell types. A cell line of RGP primary melanoma had characteristics of both benign and malignant cells: nevus-like morphology; nontumorigenicity in nude mice; but karyotypic abnormality of chromosome 6. It also had a ganglioside pattern similar to that of normal melanocytes but not melanomas, i.e., a high G_M3 ganglioside content compared to the amounts of G_M2, G_D2, and G_D3 gangliosides. Binding of monoclonal antibodies secreted by hybridomas generated by immunization of mice with VGP primary and metastatic melanoma was highest with cells and supernatants of cultures from advanced melanoma and least with nevus cells. There was no binding to normal melanocytes except with the monoclonal antibodies specific for nerve growth factor receptor or 9-O-acetyl-G_D3 ganglioside. On the other hand, monoclonal anti-nevus antibodies bound to melanocytes, nevus cells, and RGP primary melanoma cells but not to VGP primary or metastatic melanoma cells. Cultured human melanocytic cells appear to be a unique model for the study of tumor progression.

¹ This work was supported by NIH Grants CA-25874, CA-21124, CA-10815, CA-29200, and CA-25298.

² To whom requests for reprints should be addressed.

Received 4/29/85. Revised 8/ 8/85. Accepted 8/12/85.