Skip to main content
Log in

Flow cytometric analysis of the DNA content in cultured human brain tumor cells

  • Published:
Virchows Archiv B

Summary

Analysis of DNA content in cultured cells derived from 5 benign and 8 malignant human brain tumors was performed by flow cytometry, using propidium iodide as fluorochrome. Normal, non-stimulated human lymphocytes were used as controls. Cells harvested from the first confluent subculture had DNA distribution histograms similar to those of a replicating, non-synchronous population of diploid cells. This observation was made on all cultures, regardless of the DNA distribution patterns in the original tumors, including two metastatic tumors which originally had a predominantly triploid DNA content. After 3 to 5 transfers an increase in the proportion of cells with a wide range of DNA content was observed in some cultures. These changes were usually associated with a loss of cell viability. These observations suggest that in the experimental setting described, in vitro proliferation of human tumor cells may be selectively associated with cells with diploid amounts of DNA.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Atkin NB (1962) The relationship between the deoxyribonucleic acid content and ploidy in human tumors. Cytogenetics 1:113–117

    PubMed  CAS  Google Scholar 

  • Böyum A (1968) Separation of leucocytes from blood and bone marrow, with special reference to factors which influence and modify sedimentation properties of hematopoietic cells. III. A one-stage procedure for isolation of granulocytes and lymphocytes from human blood. General sedimentation properties of white blood cells in a 1 g gravity field. Scand J Clin Lab Invest 21 (Suppl 97): 51–76

    Google Scholar 

  • Cram LS, Lehman JM (1977) Flow microfluorometric DNA content measurements of tissue culture cells and peripheral lymphocytes. Hum Genet 37:201–206

    Article  PubMed  CAS  Google Scholar 

  • De Reuck J, Roels H, Vander Eecken H (1979) Cytophotometric DNA determination in human astroglial tumours. Histopathology 3:107–115

    Article  PubMed  Google Scholar 

  • Frederiksen P, Reske-Nielsen E, Bichel P (1978) Flow cytometry in tumours of the brain. Acta Neuropathol 41:179–183

    Article  PubMed  CAS  Google Scholar 

  • Frederiksen P, Reske-Nielsen E, Bichel P (1979) DNA content of meningiomas. Acta Neuropathol 46:65–68

    Article  PubMed  CAS  Google Scholar 

  • Fried J, Perez AG, Clarkson BD (1978) Rapid hypotonic method for flow cytofluorometry of monolayer cell cultures. Some pitfalls in staining and data analysis. J Histochem Cytochem 26:921–933

    PubMed  CAS  Google Scholar 

  • Fulwyler MJ (1979) Standards for flow cytometry. In: Melamed MR, Mullaney PF, Mendelsohn ML (eds) Flow cytometry and sorting. John Wiley & Sons, New York, Chichester, Brisbane, Toronto p 351–358

    Google Scholar 

  • Heller JA, Elliot KA (1954) Desoxyribonucleic acid content and cell density in brain and human brain tumors. Canad J Biochem Physiol 32:584–592

    PubMed  CAS  Google Scholar 

  • Horan, PK, Wheeles Jr LL (1977) Quantitative single cell analysis and sorting. Science 198:149–157

    Article  PubMed  CAS  Google Scholar 

  • Hoshino T, Nomura K, Wilson CB, Knebel KB, Gray JW (1978) The distribution of nuclear DNA from human brain tumor cells. J Neurosurg 49:13–21

    Article  PubMed  CAS  Google Scholar 

  • Kaji K, Matsuo M (1979) Aging of chick embryo fibroblasts in vitro. III Polyploid cell accumulation. Exp Cell Res 119:231–236

    Article  PubMed  CAS  Google Scholar 

  • Kajikawa H, Kawamoto K, Herz F, Wolley RC, Hirano A, Koss LG (1978) Flow-through cytometry of meningiomas and cultured meningioma cells. Acta Neuropathol 44:183–187

    Article  PubMed  CAS  Google Scholar 

  • Kawamoto K, Herz F, Wolley RC, Hirano A, Kajikawa H, Koss LG (1979) Flow cytometric analysis of the DNA distribution in human brain tumors. Acta Neuropathol 46:39–44

    Article  PubMed  CAS  Google Scholar 

  • Khominsky BS, Brodskaya IA (1973) Histochemistry of nucleic acids and proteins in cells of neuroectodermal tumours of different grades of malignancy. Neoplasma 20:281–296

    PubMed  CAS  Google Scholar 

  • Kraemer PM, Petersen DF, Van Dilla MA (1971) DNA constancy in heteroploidy and the stemline theory of tumors. Science 174:714–717

    Article  PubMed  CAS  Google Scholar 

  • Krishan A (1975) Rapid flow cytofluorometric analysis of mammalian cell cycle by propidium iodide staining. J Cell Biol 66:188–193

    Article  PubMed  CAS  Google Scholar 

  • Krishan A, Ganapathi RN, Israel M (1978) Effect of adriamycin and analogs on the nuclear fluorescence of propidium iodide-stained cells. Cancer Res 38:3656–3662

    PubMed  CAS  Google Scholar 

  • Lehmann J, Krug H (1980) Flow-through fluorocytophotometry of different brain tumours. Acta Neuropathol 49:123–132

    Article  PubMed  CAS  Google Scholar 

  • Melamed MR, Darzynkiewicz Z, Traganos F, Sharpless TK (1977) Nucleic acid content and nuclear chromatin structure of human bladder cell culture lines as studied by flow cytofluorometry. Cancer Res 37:1227–1231

    PubMed  CAS  Google Scholar 

  • Müller W, Brämisch R, Afra D, Schwenzfeger A (1977) Cytophotometrische Messungen des DNS-Gehaltes in Ependimomen und Plexuspapillomen. Acta Neuropathol 39:255–259

    Article  PubMed  Google Scholar 

  • Pevsner LZ, Tomina ED, Chaika TV (1964) Cytospectrophotometric study of DNA content in the cells of human brain tumours (Russian text). Vopr Med Khim 10:379–386

    Google Scholar 

  • Scarloto G, Müller W (1959) Quantitative histochemische Untersuchungen des Desoxyribonukleinsäure-Gehaltes in Hirntumoren. Zentralbl Neurochir 19:179–185

    Google Scholar 

  • Schaeffer WI (1979) Proposed usage of animal tissue culture terms (Revised 1978). Usage of vertebrate cell tissue and organ culture terminology. In Vitro 15:649–653

    Article  PubMed  CAS  Google Scholar 

  • Schneider EL, Fowlkes BJ (1976) Measurement of DNA content and cell volume in senescent human fibroblasts utilizing flow multiparameter single cell analysis. Exp Cell Res 98:298–302

    Article  PubMed  CAS  Google Scholar 

  • Suzuki N, Withers HR, Lee LY (1977) Variability of DNA content of murine fibrosarcoma cells. Nature 269:531–532

    Article  PubMed  CAS  Google Scholar 

  • Van Dilla MA, Mendelsohn ML (1979) Introduction and resumé of flow cytometry and sorting. In: Melamed MR, Mullaney PF, Mendelsohn ML (eds) Flow cytometry and sorting, John Wiley & Sons, New York, Chichester, Brisbane, Toronto p 11–37

    Google Scholar 

  • Wolley RC, Dembitzer HM, Herz F, Schreiber K, Koss LG (1976) The use of a slide spinner in the analysis of cell dispersion. J Histochem Cytochem 24:11–15

    PubMed  CAS  Google Scholar 

  • Wolley RC, Kawamoto K, Herz F, Hirano A, Koss LG (1979) Flow cytometry of human brain tumors. In: Zimmerman HM (ed) Progress in Neuropathology, vol 4. Raven Press, New York, p 267–276

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kawamoto, K., Herz, F., Wolley, R.C. et al. Flow cytometric analysis of the DNA content in cultured human brain tumor cells. Virchows Archiv B Cell Pathol 35, 11–17 (1980). https://doi.org/10.1007/BF02889145

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02889145

Key words

Navigation