Human Neoplastic and Normal Cells in Tissue Culture. I. Cell Lines Derived From Malignant Melanomas and Normal Melanocytes2

Abstract

Forty-six cell lines derived from 31 human melanomas obtained from 28 patients were cultured. Fourteen of 16 lines have produced malignant tumors when injected into nude (thymus-deficient) mice. Tumors in 5 of the nude mice metastasized to distant lymph nodes and/or to the lungs of the mouse host. Extreme variability from line to line was observed for doubling time (34 to 106 hr), plating efficiency (0-86%), and melanin production. All tested lines had type B glucose-6-phosphate dehydrogenase, thereby excluding HeLa cell contamination. HeLa cells have been grown for some time in our laboratory. Our results clearly demonstrated that HeLa cell contamination does not occur invariably in heteroploid lines growing in a laboratory simultaneously with Hela cells, provided that proper care is taken to avoid such occurrence. Multiple cell lines derived from the same tumor had identical phosphoglucomutase enzyme phenotype, which suggested a lack of significant cross-contamination between the lines. Four long-term cultures of normal human uveal embryo melanocytes have also been established and characterized. Although all produced melanin after reaching saturation density, they differed from the melanoma cells morphologically; they were flat, not refringent, and lacked piling up and plating ability. When melanoma cells were exposed to bromodeoxyuridine (BUDR) for long periods, a phenotypic change toward non-neoplastic characteristics was observed. Cells became flat and not refringent and, when injected into nude mice, tumors appeared after a long latent period. These changes were completely reversible in vitro and in vivo. The BUDR-treated cultures were undistinguishable from the untreated mother cultures after 2 to 3 passages. Lines derived from tumors in nude mice (obtained by injection of BUDR-treated cells) were again indistinguishable from the untreated mother line. Normal melanocytes were mostly euploid; all the melanoma cells were aneuploid. All 29 cell lines derived from 14 patients had an average chromosome number higher than 46. Detailed group-by-group chromosome analysis always showed an excess of C chromosomes, which suggested that hyperreduplication of one or more C chromosomes is a specific characteristic of human melanomas.

Characterization of seven human melanoma cell lines: Melanogenesis and secretion of plasminogen activators

“…The two cell lines in this series that did pigment in culture were similar to those described by Romsdahl and Hsu (1972) and Giovanella et al (1976) in that melanogenesis increased with increasing cell density. In our experience cell clusters that formed at confluence were particularly deeply pigmented.…”

The cancer growth suppressing gene mda-7 induces apoptosis selectively in human melanoma cells

“…HO-1 and MeWo are established human melanotic melanoma cell lines and FO-1 is an established human amelanotic melanoma cell line isolated from separate patient-derived metastatic lesions (Giovanella et al, 1976;Kerbel et al, 1994;Fisher et al, 1985;Graham et al, 1991). WM35 was derived from an RGP primary human melanoma (Herlyn, 1990).…”

Suppression of human ribosomal protein L23A expression during cell growth inhibition by interferon-β

“…The HO-1 human melanoma cell line was established from a metastatic inguinal lymph node lesion from a 49 year-old female (Giovanella et al, 1976;. HeLa is a human cervical carcinoma cell line (Kao and Nevins, 1983).…”