Abstract
BackgroundMalignant mesothelioma is an aggressive tumour of serosal surfaces most commonly pleura. Characterised cell lines represent a valuable tool to study the biology of mesothelioma. The aim of this study was to develop and biologically characterise six malignant mesothelioma cell lines to evaluate their potential as models of human malignant mesothelioma.MethodsFive lines were initiated from pleural biopsies, and one from pleural effusion of patients with histologically proven malignant mesothelioma. Mesothelial origin was assessed by standard morphology, Transmission Electron Microscopy (TEM) and immunocytochemistry. Growth characteristics were assayed using population doubling times. Spectral karyotyping was performed to assess chromosomal abnormalities. Authentication of donor specific derivation was undertaken by DNA fingerprinting using a panel of SNPs.ResultsMost of cell lines exhibited spindle cell shape, with some retaining stellate shapes. At passage 2 to 6 all lines stained positively for calretinin and cytokeratin 19, and demonstrated capacity for anchorage-independent growth. At passage 4 to 16, doubling times ranged from 30–72 hours, and on spectral karyotyping all lines exhibited numerical chromosomal abnormalities ranging from 41 to 113. Monosomy of chromosomes 8, 14, 22 or 17 was observed in three lines. One line displayed four different karyotypes at passage 8, but only one karyotype at passage 42, and another displayed polyploidy at passage 40 which was not present at early passages. At passages 5–17, TEM showed characteristic features of mesothelioma ultrastructure in all lines including microvilli and tight intercellular junctions.ConclusionThese six cell lines exhibit varying cell morphology, a range of doubling times, and show diverse passage-dependent structural chromosomal changes observed in malignant tumours. However they retain characteristic immunocytochemical protein expression profiles of mesothelioma during maintenance in artificial culture systems. These characteristics support their potential as in vitro model systems for studying cellular, molecular and genetic aspects of mesothelioma.
Highlights
Malignant mesothelioma is an aggressive tumour of serosal surfaces, usually caused by exposure to asbestos
Because of long latency from exposure to diagnosis, the incidence of mesothelioma is expected to increase in parts of the world where asbestos was mined or where asbestos products were used
This study aimed 1) to establish a bio-tool of annotated human malignant mesothelioma tumour cell lines characterised for morphology, doubling times, immunocytochemical properties, and chromosomal alterations, and 2) to define passage related changes in cytogenetic profiles of human mesothelioma cell lines in long term culture
Summary
Malignant mesothelioma is an aggressive tumour of serosal surfaces (most commonly pleura), usually caused by exposure to asbestos. Better understanding of the biology of mesothelioma underpins discovery and implementation of new therapeutic strategies. To this end, in vitro models of disease have been important tools for studying biological properties of tumours. Malignant mesothelioma is an aggressive tumour of serosal surfaces most commonly pleura. Characterised cell lines represent a valuable tool to study the biology of mesothelioma. The aim of this study was to develop and biologically characterise six malignant mesothelioma cell lines to evaluate their potential as models of human malignant mesothelioma
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