Establishment and characterization of five human malignant mesothelioma cell lines derived from pleural effusions

Our plain-English summary of Establishment and characterization of five human malignant mesothelioma cell lines derived from pleural effusions, by L.S. Manning, D. Whitaker, A.R. Murch, et al (International Journal of Cancer, 1991).

The study

• Authors: L.S. Manning, D. Whitaker, A.R. Murch, M.J. Garlepp, M.R. Davis, A.W. Musk, B.W. Robinson
• Journal: International Journal of Cancer
• Published: 1991;47(2):285-90
• Cited by: 85 articles, as counted by NIH iCite
• Read it: PubMed · Publisher

Abstract

Malignant mesothelioma (MM) is an aggressive tumour of the serosal cavities which is associated with exposure to asbestos. Studies of this tumour have been limited by a paucity of well-characterized human MM cell lines. In this study, 5 human MM cell lines were established from pleural effusions of patients with this malignancy. All 5 patients were males with known crocidolite asbestos exposure, who had received no treatment for their disease and in whom the diagnosis was confirmed by cytology, histology and electron microscopy (EM). These lines have been in culture from 11 to 25 months, and all of them for more than 18 passages. The appearance of the cells in culture was extremely varied; in 3 of the lines they were spindle-shaped with few vacuoles (JU77, LO68 and ONE58); in 1 line they had a thick, stellate shape with vacuoles (NO36) and in 1 they were very pleomorphic in both shape and size with irregular membranes and numerous vacuoles [DeH128 (M)]. Upon reaching confluence, cells in 3 of the 5 lines assumed the cobblestone-like pattern characteristic of epithelial-type cells, whereas in the other 2 (LO68 and ONE58) they remained spindle-shaped. All 5 lines demonstrated a loss of contact inhibition (i.e., piling) at confluence. Minimum doubling times varied significantly from 18 hr (JU77) to more than 30 hr [DeH128 (M)]. Cytological examination showed characteristic mesothelial/mesothelioma morphology, and epithelial membrane antigen (EMA) and cytokeratin were demonstrated in cells from all 5 lines. These cells lacked CEA and epithelial mucin. The presence of cell junctions, glycogen and numerous long, thin, branching microvilli was readily demonstrable by EM. All lines had abnormal karyotypes, with the modal chromosome number varying from 40 to 80. Variable chromosome numbers, numerous structural rearrangements and unrecognizable marker chromosomes were readily observed; however, the only consistent change seen was del 6q21 in 4 of the 5 lines. The establishment of these 5 cultured human MM cell lines now provides an opportunity for comparative study of several aspects of the biology of MM in vitro as well as screening new treatment modalities.

In plain English

This Western Australian laboratory study described five malignant mesothelioma cell lines grown from pleural fluid of five men aged 36 to 62. All five had pleural mesothelioma and known crocidolite asbestos exposure. The lines had been grown for at least 11 months and at least 18 passages. Under standard conditions, average doubling times ranged from 25 hours (JU77) to 46 hours (DeH128 (M)). All five lines showed the markers EMA and cytokeratin and lacked epithelial mucin; only NO36 had a small amount of CEA. The authors wrote that over 90% of cells in four lines reacted with two anti-mesothelial antibodies, but their table shows this only for MM-2; MM-1 was reported for three lines (94.3%, 92.7%, 88.3%) and not done for two. DeH128 (M), which had been passed through nude mice, reacted less. All lines had abnormal chromosomes, with modal numbers from 40 to 80, and a deletion at 6q21 appeared in 4 of the 5 lines. The authors concluded that no consistent chromosome change characteristic of mesothelioma was found, and that the lines could help screen treatments.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: Laboratory study establishing and characterizing human mesothelioma cell lines grown in culture
• Who was studied: Cells from pleural effusions of 5 male patients aged 36 to 62 with pleural malignant mesothelioma, at Sir Charles Gairdner Hospital and the Queen Elizabeth II Medical Centre, Nedlands, Western Australia. The tumours were subtyped as epithelial from the effusion samples.
• Exposure: Known crocidolite asbestos exposure in all 5 patients. No doses or jobs are given. The authors relate the local disease rate to crocidolite mining at Wittenoom Gorge, 1943 to 1966.
• Main results:
  • The lines had been in culture from 11 to 25 months, for at least 18 passages.
  • Under standard culture conditions, average doubling times ranged from 25 hours (JU77) to 46 hours (DeH128 (M)).
  • Under conditions set to find minimum doubling times, these fell to 18 hours (JU77) and 30 hours (DeH128 (M)).
  • EMA and cytokeratin were found in all 5 lines.
  • A small amount of CEA was found in line NO36 only. All 5 lines were negative for epithelial mucin.
  • In the text, the authors stated that more than 90% of cells from 4 of the 5 lines stained with 2 mouse anti-human mesothelial antibodies.
  • In Table I, the MM-2 antibody stained 93.4%, 92.9%, 90.5% and 96.3% of cells in four lines, and 24.7% in a fifth (apparently DeH128 (M)).
  • In Table I, the MM-1 antibody was reported for three lines (94.3%, 92.7% and 88.3% of cells) and marked not done (ND) for the remaining two.
  • DeH128 (M), established after passage through nude mice, showed significantly less reactivity with these antibodies.
  • All 5 lines had abnormal karyotypes, with modal chromosome numbers from 40 to 80.
  • A deletion of the long arm of chromosome 6, with a breakpoint at 6q21, was seen in 4 of the 5 lines. No abnormality was common to all lines.
  • Testing confirmed that each line was genetically unique (no cross-contamination).
  • Samples from 11 of 33 patients with mesothelioma were established in culture for more than 3 months, which the authors give as a 33% establishment rate.
  • Two lines, DeH128 (M) and ONE58, formed tumours in nude mice, and xenograft models were established for both.
• The authors' conclusion: The authors concluded that no consistent chromosome aberration characteristic of mesothelioma had been found. They also concluded that the panel of lines could be used in laboratory screening that may help identify treatments to try in patients.
• Limitations the authors state:
  • In three lines (NO36, JU77, ONE58), many unrecognizable marker chromosomes made it impossible to determine their true chromosomal makeup accurately.
  • Of the lines established, only these 5 had been fully characterized.

Topics

The National Library of Medicine indexes this study under subjects our site covers: Mesothelioma, Pleural plaques and thickening.

Medical terms in this study, explained in our Lexicon: Malignant Mesothelioma, Pleural Effusion, Crocidolite, Asbestos Exposure, Cytology, Histology, Pleomorphic, Mesothelioma.

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