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Establishment and characterization of an uveal-melanoma cell line

✍ Scribed by Itte De Waard-Siebinga; Derk-Jan R. Blom; Marieke Griffioen; Peter I. Schrier; Ed Hoogendoorn; Geoff Beverstock; Erik H. J. Danen; Marline J. Jager

Publisher
John Wiley and Sons
Year
1995
Tongue
French
Weight
996 KB
Volume
62
Category
Article
ISSN
0020-7136

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✦ Synopsis

Abstract

A human uveal melanoma cell line (92–1) was established from a primary uveal melanoma, and has now been maintained in culture for over 2½ years. Light microscopy of the cultured cells demonstrated extremely pleiomorphic cells with large prominent nucleoli. Cell proliferation was determined with a non‐radioactive propidium‐iodide assay and indicated an in vitro doubling time of approximately 58 hr. Furthermore, the cell line was characterized by cytogenetic analysis, electron microscopy, immunocytochemistry and Northern blotting for HLA and c‐myc‐mRNA analysis. Cytogenetic analysis revealed numerical abnormalities of chromosome 8 and structural abnormalities of chromosome 6. By electron microscopy, different stages of melanosome development were observed. Immunocytochemical analysis demonstrated expression of the melanoma‐associated antigen gp100. Expression analysis of HLA antigens revealed a very low level of, in particular, the HLA‐B locus products, which could be induced by interferon‐α or ‐γ treatment. Likewise, Northern‐blot experiments revealed decreased levels of HLA‐B mRNA as compared with HLA‐A. In addition, high levels of c‐myc expression were observed. The phenotypic characteristics of the cultured cells indicate that we have established an uveal melanoma cell line. This now well‐characterized uveal melanoma cell line can be used in future studies. © 1995 Wiley‐Liss, Inc.

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