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Pharmaceutisch Weekblad

, Volume 11, Issue 5, pp 155–160 | Cite as

Rapid assay for thein vitro chemosensitivity testing of human breast tumours

Control of assay conditions and quality assurance
  • R. F. M. Oude Elferink
  • H. M. J. Goldschmidt
  • H. J. Majoor
  • J. F. Leijten
Original Articles

Abstract

This study has been initiated by the definite need of a rapid,in vitro and patient-specific test to examine oncolytic drug effects on tumour cells. Therefore, we applied a test in which we monitored the incorporation of labelled nucleosides in isolated tumour cells, as a measure of nucleus activity (i.e. the Volm assay). A novel technique of erythrocytes co-precipitation has been developed and this enabled us to use a small number of tumour cells per test (200,000 cells/tube). During the assay, a strict pH control and a high starting viability have been introduced. A cytotoxic control and a t-test at two levels deal with the technical errors and the imprecision. For the evaluation of a specific drug a number of 1.8 million cells proved to be sufficient. Time-course studies of the incorporation of labelled nucleosides into isolated tumour cells have shown the optimal incubation time to be 2 h. The entire assay is completed within one day. HeLa cell cultures were employed as quality control material and criteria for interpretation have been developed. Preliminary results, based upon the evaluation of 43 human breast tumours with doxorubicin, indicate the correctness of the proposed procedures. In conclusion we can state that this assay not only provides useful information but above all that the results are made available in such a short time that they can be used directly in the medical management of the individual patient.

Keywords

Antineoplastic agents Breast neoplasms Doxorubicin Drug resistance Quality control Sensitivity tests,in vitro 

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References

  1. 1.
    Mattern J, Volm M. Clinical relevance of predictive tests for cancer chemotherapy. Cancer Treat Rev 1982;9: 267–98.PubMedGoogle Scholar
  2. 2.
    Weisenthal LM, Lippman ME. Clonogenic and nonclonogenicin vitro chemosensitivity assays. Cancer Treat Rep 1985;69:615–32.PubMedGoogle Scholar
  3. 3.
    Hamburger AW, Salmon SE. Primary bioassay of human tumor stem cells. Science 1977;197:461–3.PubMedGoogle Scholar
  4. 4.
    Volm M, Wayss K, Kaufmann M, Mattern J. Pretherapeutic detection of tumour resistance and the results of tumour chemotherapy. Eur J Cancer 1979;15:983–93.PubMedGoogle Scholar
  5. 5.
    Wilson AP, Tayler C, Laher B, Newman C, Ford C, Howell A. Results obtained in short-term assay using nucleotide incorporation for thein vitro prediction of chemosensitivity of ovarian tumors [Abstract]. Br J Cancer 1983;48:119.Google Scholar
  6. 6.
    Group for Sensitivity Testing of Tumours. KSST.In vitro short-term test to determine the resistance of human tumors to chemotherapy. Cancer 1981;48: 2127–35.Google Scholar
  7. 7.
    Sokal RR, Rohlf FJ. Biometry. The principles and practice of statistics in biological research. 2nd ed. New York: W.H. Freeman and Company, 1981:429.Google Scholar
  8. 8.
    Oude Elferink RFM, Spaan LWJ, Majoor HJ, Goldschmidt HMF. Quality assessment in short-term cytostatic drug testing of human tumor cells [Abstract]. J Clin Chem Clin Biochem 1988;26:318.Google Scholar
  9. 9.
    Kern DH, Drogemuller CR, Kennedy MC, Hildebrand-Zanki SU, Tanigawa N, Sondak VK. Development of a miniaturized, improved nucleic acid precursor incorporation assay for chemosensitivity testing of human solid tumors. Cancer Res 1985;45:5436–41.PubMedGoogle Scholar
  10. 10.
    Dittrich CH, Jakesz R, Wrba F, et al. The human tumor cloning assay in the management of breast cancer patients. Br J Cancer 1985;52:197–203.PubMedGoogle Scholar
  11. 11.
    Harris Jr, Hellman S, Canellos GP, Fisher F. Cancer of the Breast. In: DeVita VT Jr, Hellman S, Rosenberg SA, eds. Cancer. Principles & practice of oncology. 2nd ed. Philadelphia: Lippincott, 1985:1119–77.Google Scholar
  12. 12.
    Wilson AP, Ford CHJ, Newman CE, Howell A. A comparison of three assays used for thein vitro chemosensitivity testing of human tumors. Br J Cancer1984;49: 57–63.PubMedGoogle Scholar
  13. 13.
    Slocum HK, Pavelic ZP, Rustum YM, et al. Characterization of cells obtained by mechanical and enzymatic means from human melanoma, sarcoma, and lung tumors. Cancer Res 1981;41:1428–34.PubMedGoogle Scholar
  14. 14.
    Good NE, Winget GD, Winter W, Connolly TN, Izawa S, Singh RMM. Hydrogen ion buffers for biological research. Biochemistry 1966;5:467–77.PubMedGoogle Scholar
  15. 15.
    Tanigawa N, Kern DH, Hikasa Y, Morton DL. Rapid assay for evaluating the chemosensitivity of human tumors in soft agar culture. Cancer Res 1982;42: 2159–64.PubMedGoogle Scholar

Copyright information

© Royal Dutch Association for Advancement of Pharmacy 1989

Authors and Affiliations

  • R. F. M. Oude Elferink
    • 1
  • H. M. J. Goldschmidt
    • 2
  • H. J. Majoor
    • 2
  • J. F. Leijten
    • 1
  1. 1.Department of Clinical Chemistry and HaematologySt. Elisabeth HospitalLC Tilburgthe Netherlands
  2. 2.Department of Laboratory Oncology and HaematologyDr. Bernard Verbeeten InstituteLA Tilburgthe Netherlands

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