Advertisement

Three-dimensional growth patterns of various human tumor cell lines in simulated microgravity of a NASA bioreactor

  • M. Ingram
  • G. B. Techy
  • R. Saroufeem
  • O. Yazan
  • K. S. Narayan
  • T. J. Goodwin
  • G. F. Spaulding
Cellular Models

Summary

Growth patterns of a number of human tumor cell lines that form three-dimensional structures of various architectures when cultured without carrier beads in a NASA rotary cell culture system are described and illustrated. The culture system, which was designed to mimic microgravity, maintained cells in suspension under very low-shear stress throughout culture. Spheroid (particulate) production occurred within a few hours after culture was started, and spheroids increased in size by cell division and fusion of small spheroids, usually stabilizing at a spheroid diameter of about 0.5 mm. Architecture of spheroids varied with cell type. Cellular interactions that occurred in spheroids resulted in conformation and shape changes of cells, and some cell lines produced complex, epithelial-like architectures. Expression of the cell adhesion molecules, CD44 and E cadherin, was upregulated in the three-dimensional constructs. Coculture of fibroblast spheroids with PC3 prostate cancer cells induced tenascin expression by the fibroblasts underlying the adherent prostate epithelial cells. Invasion of the fibroblast spheroids by the malignant epithelium was also demonstrated.

Key words

spheroid architecture suspension culture low-shear culture 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Acker, H. Microenvironmental conditions in multicellular spheroids grown over liquid-overlay tissue culture conditions. In: Recent results in cancer research. Vol. 95. Berlin, Heidelberg: Springer-Verlag; 1984:116–133.Google Scholar
  2. 2.
    Bauer, K. D.; Keng, P.; Sutherland, R. M. Isolation of quiescent cells from multicellular tumor spheroids using centrifugal elutriation. Cancer Res. 42:3956–3965; 1980.Google Scholar
  3. 3.
    Becker, J. L.; Prewett, T. L.; Spaulding, G. F., et al. Three-dimensional growth and differentiation of ovarian tumor cell line in high aspect rotating wall vessel: morphologic and embryologic considerations. J. Cell. Biochem. 51:283–290; 1993.PubMedCrossRefGoogle Scholar
  4. 4.
    Carlsson, J. A proliferation gradient in three-dimensional colonies of cultured human glioma cells. Int. J. Cancer 20:129–136; 1977.PubMedCrossRefGoogle Scholar
  5. 5.
    Dall, P.; Heider, K.-H.; Sinn, H.-P., et al. Comparison of immunohistochemistry and RT/PCR for detection of CD44v-expression, a new prognostic factor in human breast cancer. Int. J. Cancer 60:471–477; 1995.PubMedCrossRefGoogle Scholar
  6. 6.
    Erickson, H. P.; Bourdon, M. A. Tenascin: an extracellular matrix protein prominent in specialized embryonic tissues and tumors. Ann. Rev. Cell Biol. 5:71–92; 1989.PubMedGoogle Scholar
  7. 7.
    Fox, S. B.; Gotter, K. C.; Jackson, D. G., et al. CD44 and cancer screening. Lancet 342:548–549; 1993.PubMedCrossRefGoogle Scholar
  8. 8.
    Freyer, J. P.; Sutherland, R. M. Selective dissociation and characterization of cells from different regions of multicell tumor spheroids. Cancer Res. 40:3956–3965; 1980.PubMedGoogle Scholar
  9. 9.
    Goodwin, T. J.; Jessup, J. M.; Wolf, D. A. Morphologic differentiation of colon carcinoma cell lines HT-29 and HT-29KM in rotating wall vessels. In Vitro Cell. Dev. Biol. 28A:47–60; 1992.PubMedCrossRefGoogle Scholar
  10. 10.
    Goodwin, T. J.; Schroeder, W. F.; Wolf, D. A., et al. Rotating-wall vessel coculture of small intestine as a prelude to tissue modeling: aspects of simulated microgravity. Proc. Soc. Exp. Biol. Med. 202:181–192; 1993.PubMedGoogle Scholar
  11. 11.
    Ingram, M.; Buckwalter, J. G.; Jacques, D. B., et al. Immunotherapy for recurrent malignant glioma: an interim report on survival. Neurol. Res. 12:265–273; 1990.PubMedGoogle Scholar
  12. 12.
    Kaighn, M. E.; Narayan, K. S.; Ohnuki, Y., et al. Establishment and characterization of a human prostatic carcinoma cell line (PC-3). Invest. Urol. 17:16–23; 1979.PubMedGoogle Scholar
  13. 13.
    Kato, K.; Hibino, S.; Yagita, H., et al. Tenascin suppresses T-cell activation mediated by CD3 and costimulation molecules. Abs. #5085, 9th International Congress of Immunology, 23–29 July 1995, San Francisco, CA.Google Scholar
  14. 14.
    Laerum, O. D.; Bjerkrig, R. Monolayer and three-dimensional culture of rat and human central nervous system: normal and malignant cells and their interactions. Methods in neurosciences. Vol. 2. London: Academic Press; 1990:210–236.Google Scholar
  15. 15.
    Matsumura, T.; Tarin, D. Significance of CD44 gene products for cancer diagnosis and disease evaluations. Lancet 340:1053–1058; 1992.PubMedCrossRefGoogle Scholar
  16. 16.
    Mueller-Lieser, W. Multicellular spheroids. J. Cancer Res. Clin. Oncol. 13:101–122; 1987.CrossRefGoogle Scholar
  17. 17.
    Overduin, M.; Harvey, T. S.; Bagby, S., et al. Solution structure of the epithelial cadherin domain responsible for selective cell adhesion. Science 267:386–389; 1995.PubMedCrossRefGoogle Scholar
  18. 18.
    Prewett, T. L.; Goodwin, T. J.; Spaulding, G. F. Three-dimensional modeling of T-24 human bladder carcinoma cell line: a new simulated microgravity culture vessel. J. Tissue Culture Methods 15:29–36; 1993.CrossRefGoogle Scholar
  19. 19.
    Rutgers, D. H.; Dorien, P. P. N.; van der Linden, P. M. Cell kinetics of hypoxic cells in a murine tumor in vivo: flow cytometric determination of the radiation-induced blockage of cell cycle progression. Cell Tissue Kinet. 20:37–42; 1987.PubMedGoogle Scholar
  20. 20.
    Schwarz, R. P.; Goodwin, T. J.; Wolf, D. A. Cell culture for three-dimensional modeling in rotating wall-vessels: an application of simulated microgravity. J. Tissue Culture Methods 14:51–58; 1992.CrossRefGoogle Scholar
  21. 21.
    Stamenkovic, I.; Aruffo, A.; Amiot, M., et al. The hematopoietic and epithelial forms of CD44 are distinct polypeptides with different adhesion potentials for hyaluronate-bearing cells. EMBO 10:343–348; 1991.Google Scholar
  22. 22.
    Sutherland, R. M. Cell and microenvironment in tumor microregions. Science 240:177–184; 1988.PubMedCrossRefGoogle Scholar
  23. 23.
    Sutherland, R. M.; McCredie, I. A.; Inch, W. R. Growth of multicellular spheroids in tissue culture as a model of nodular carcinoma. J. Natl. Cancer Inst. 46:113; 1971.PubMedGoogle Scholar
  24. 24.
    Sutherland, R. M.; Sordat, B.; Bamat, J., et al. Oxygenation and differentiation in multicellular spheroids of human colon carcinoma. Cancer Res. 46:5320–5329; 1986.PubMedGoogle Scholar
  25. 25.
    Yuhas, J. M.; Li, A. P.; Martinez, A. G., et al. A simplified method for production and growth of multicellular tumor spheroids. Cancer Res. 37:3639–3643; 1977.PubMedGoogle Scholar

Copyright information

© Society for In Vitro Biology 1997

Authors and Affiliations

  • M. Ingram
    • 1
  • G. B. Techy
    • 1
  • R. Saroufeem
    • 1
  • O. Yazan
    • 1
  • K. S. Narayan
    • 1
  • T. J. Goodwin
    • 2
  • G. F. Spaulding
    • 3
  1. 1.Huntington Medical Research InstitutesPasadena
  2. 2.L. B. Johnson Space CenterHouston
  3. 3.Clear Lake Medical Foundation, Inc.Houston

Personalised recommendations