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A comprehensive characterization of pancreatic ductal carcinoma cell lines: towards the establishment of an in vitro research platform

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Abstract

There are a large number of stable pancreatic ductal carcinoma cell lines that are used by researchers worldwide. Detailed data about their differentiation status and growth features are, however, often lacking. We therefore attempted to classify commonly used pancreatic carcinoma cell lines according to defined cell biological criteria. Twelve pancreatic ductal adenocarcinoma cell lines were cultured as monolayers and spheroids and graded according to their ultrastructural features. The grading system was based on the integrity of membrane structures and on the presence of mucin granules, cell organelles, nuclear and cellular polymorphism, cell polarity, and lumen formation. On the basis of the resulting scores the cell lines were classified as well, moderately, or poorly differentiated. In addition, immunocytochemistry was performed for the markers cytokeratin 7, 8, 18, 19, carcinoembryonic antigen, MUC1 MUC2, MUC5, and MUC6. The population doubling time of monolayer cultures, determined by a tetrazolium salt based proliferation assay was correlated with the ultrastructural grade. The grading of the ultrastructural features of the monolayers, and particularly of the spheroids, revealed that Capan-1 and Capan-2 cells were well differentiated; Colo357, HPAF-2, Aspc-1, A818-4, BxPc3, and Panc89 cells were moderately differentiated and PancTu-I, Panc1, Pt45P1, and MiaPaCa-2 cells poorly differentiated. Membrane-bound MUC1 staining was a characteristic of well differentiated cell lines. The population doubling time of the monolayer cultures was related to the differentiation grade. No relationship was found between the p53, K-ras, DPC4/Smad4, or p16INK4a mutation status and the grade of differentiation. We conclude that the proposed ultrastructural grading system combined with the proliferative activity provides a basis for further comparative studies of pancreatic ductal adenocarcinoma cell lines.

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Acknowledgements

The authors thank O. Bracker, R Köpke, B. Facompré, and A. Höltzl for their skillful technical assistance, Prof. Lüllmann-Rauch and Prof. A. Kadar for their helpful technical suggestions, and K. Dege for editing the manuscript. This work was supported by the Werner and Klara Kreitz Foundation, the IZKF, University of Kiel, and a Faculty Grant.

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Authors and Affiliations

  1. Department of Pathology, University of Kiel, Michaelisstrasse 11, 24105, Kiel, Germany

    Bence Sipos, Simone Möser & Günter Klöppel

  2. Molecular Oncology Unit, Department of Abdominal and Thoracic Surgery, Michaelisstrasse 11, 24105, Kiel, Germany

    Holger Kalthoff

  3. Second Department of Pathology, Semmelweis University, Budapest, Hungary

    Virag Török

  4. Second Department of Medicine, University of Heidelberg-Mannheim, Mannheim, Germany

    Matthias Löhr

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  1. Bence Sipos
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  2. Simone Möser
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  3. Holger Kalthoff
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  4. Virag Török
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  6. Günter Klöppel
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Correspondence to Bence Sipos.

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Sipos, B., Möser, S., Kalthoff, H. et al. A comprehensive characterization of pancreatic ductal carcinoma cell lines: towards the establishment of an in vitro research platform. Virchows Arch 442, 444–452 (2003). https://doi.org/10.1007/s00428-003-0784-4

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  • DOI: https://doi.org/10.1007/s00428-003-0784-4

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