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C.J. Clark



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    P3.02 - Biology/Pathology (ID 620)

    • Event: WCLC 2017
    • Type: Poster Session with Presenters Present
    • Track: Biology/Pathology
    • Presentations: 1
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      P3.02-078 - Establishing Malignant Pleural Mesothelioma Primary Cell Lines Using the 3D Spheroid Method Produces a Model with Better Tumour Architecture (ID 10456)

      09:30 - 09:30  |  Author(s): C.J. Clark

      • Abstract
      • Slides

      Background:
      Malignant pleural mesothelioma (MPM) is an aggressive malignancy with no effective treatment options. Poor prognosis and drug resistance are the main challenges of this deadly disease. There is also no simple distinctive diagnosis tool for identification of MPM. Better diagnostic markers may also provide better biological information for newer treatment option development. In this study we have established primary MPM cell lines and characterised them with current biomarkers. Our ultimately goal is to use these cell lines for better identification of diagnostic biomarkers.

      Method:
      MPM cell lines were either established from tissue specimens or pleural effusion from patients with pathologically confirmed MPM. Cells were cultured in standard (2D) and spheroid (3D) versions for characterisation. Cells prepared in 2D and 3D were stained with H&E and analysed with a diagnostic biomarker panel (CK-8/18, Calretinin, CK5/6, CD141, WT-1, D2-40, EMA, CEA, Tag-72, BG8, CD15, TTF-1 and BAP1). Scoring and comments were provided by pathologists experienced in MPM diagnosis (KL, SK). Established cell lines were also analysed for ploidy (flow cytometry) and interphase (fluorescent microscope) for chromosome number. PBMC from healthy donor was used as a control diploid.

      Result:
      We successfully established nine cell lines from MPM patient specimens. The original tumour histological sub-types were: three epithelioid, four biphasic, one desmoplastic and one not otherwise specified. Cells grown in 3D with H&E staining revealed better tumour architecture, cell-cell contacts and morphology when compared to cells grown in standard 2D culture. Mesothelioma positive markers were more distinctive and intense in biphasic cell lines grown 3D culture. Other sub-types showed similar staining when grown in both formats. Results from ploidy showed no distinctive difference between sub-types, however, 5 out of 9 cell lines established had tetraploid chromosome content.

      Conclusion:
      Cells grown in 3D provide more tumour architecture when compared with 2D cells. 3D cells also provide more intensity and greater percentage of positive MPM markers

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