Diagnosis and Pathological Analysis of Patient Cancers by Detection of Proliferating Cells in Gelfoam® Histoculture

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1760)

Abstract

Patient tumors grew in Gelfoam® histoculture with maintenance of tissue architecture, tumor-stromal interaction, and differentiated functions. In this chapter, we review the use of Gelfoam® histoculture to demonstrate proliferation indices of major solid cancer types explanted directly from surgery. Cell proliferation was visualized by histological autoradiography within the cultured tissues after [3H]thymidine incorporation by the proliferating cells. Epilumination polarization microscopy enables high-resolution imaging of the autoradiography of each cell. The histological status of the cultured tissues can be assessed simultaneously with the proliferation status. Carcinomas were observed to have areas of high epithelial proliferation with quiescent stromal cells. Sarcomas have high proliferation of the cancer cells of mesenchymal organ. Normal tissues can also proliferate at high rates. Mean growth fraction index (GFI) was highest for patient tumors with the pure subtype of small-cell lung cancer than other types of lung cancer.

Key words

Gelfoam® Three-dimensional histoculture Tumors Lung cancer Small cell Adenocarcinoma Squamous-cell carcinoma Autoradiography Polarization Microscopy Proliferation index 

References

  1. 1.
    Freeman AE, Hoffman RA (1986) In vivo-like growth of human tumors in vitro. Proc Natl Acad Sci U S A 83:2694–2698CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Vescio RA, Redfern CA, Nelson TJ, Ugoretz S, Stern PH, Hoffman RM (1987) In vivo-like drug responses of human tumors growing in three-dimensional gel-supported primary culture. Proc Natl Acad Sci U S A 84:5029–5033CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Minna JD, Higgins GA, Glatstein EJ (1981) Cancer of the lung. In: DeVita VT, Hellman S, Rosenberg SA (eds) Principles and Practice of Oncology. J. B. Lippincott Co., Philadelphia, pp 396–474Google Scholar
  4. 4.
    Baylin SB, Weisburger WR, Eggleston JC, Mendelsohn G, Beaven M, Abeloff M, Ettinger D (1978) Variable content of histaminase. i.-dopa decarboxylase and calcitonin in small cell carcinoma of the lung. N Engl J Med 299:105–110CrossRefPubMedGoogle Scholar
  5. 5.
    Feinstem AR, Gelfman NA, Yesner R, Auerbach O, Hackel DB, Pratt PC (1970) Observer variability in the histopathologic diagnosis of lung cancer. Am Rev Respir Dis 101:671–684Google Scholar
  6. 6.
    Hirsch FR, Matthews MJ, Aisner S, Campobasso O, Elema JD, Gazdar AF, Mackay B, Nasiell M, Shimosato Y, Steele RH, Yesner R, Zettergren L (1988) Histopathologic classification of small cell lung cancer. Cancer (Phila) 2:973–977CrossRefGoogle Scholar
  7. 7.
    Vescio RA, Connors KM, Bordin GM, Robb JA, Youngkin T, Umbreit JN, Hoffman RM (1990) The distinction of small cell and non-small cell cancer by growth in native-state histoculture. Cancer Res 50:6095–6099PubMedGoogle Scholar
  8. 8.
    Hoffman RM, Monosov AZ, Connors KM, Herrera H, Price JH (1989) A general native-state method for determination of proliferation capacity of human normal and tumor tissues in vitro. Proc Natl Acad Sci U S A 86:2013–2017CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Yano S, Miwa S, Mii S, Hiroshima Y, Uehara F, Yamamoto M, Kishimoto H, Tazawa H, Bouvet M, Fujiwara T, Hoffman RM (2014) Invading cancer cells are predominantly in G0/G1 resulting in chemoresistance demonstrated by real-time FUCCI imaging. Cell Cycle 13:953–960Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.AntiCancer Inc.San DiegoUSA
  2. 2.Department of SurgeryUniversity of CaliforniaSan DiegoUSA

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