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Standardized kinetic microassay to quantify differential chemosensitivity on the basis of proliferative activity

  • Günther Bernhardt
  • Herta Reile
  • Herbert Birnböck
  • Thilo Spruß
  • Helmut Schönenberger
Original Papers Experimental Oncology

Summary

Conventionally in vitro cytotoxicity assays are performed as single-end-point determinations. To compensate for the diversity of growth rates among different cell lines in this report we describe a computerized kinetic chemosensitivity assay based on quantification of biomass by staining cells with crystal violet. As a prerequisite four human breast cancer cell line (MDA-MB-231, MCF-7, T-47-D and ZR-75-1) were characterized with regard to oestrogen and progesterone receptor content, modal chromosome number and proliferation kinetics depending on the number of passages in culture. With prolonged time in culture for ZR-75-1 exposed to various concentrations of cisplatinum a dose-related increase in drug effect was observed. Owing to a correction of the T/C values for the initial cell mass (at the time when drug is added) a sharp distinction between cytostatic and cytocidal drug effects becomes obvious in plots of corrected T/C values versus time of incubation. The influence of the untreated control on the corrected T/C values and possible time courses of theoretical inhibition profiles (reflecting cytostatic, transient cytotoxic or cytocidal drug effects as well as development of resistance) and their relationship to the corresponding growth curves of drugtreated cells are discussed. Chemosensitivity assays with diethylstilbestrol dipropionate, tamoxifen, melphalan, cisplatinum, vinblastine, Adriamycin and 5-fluorouracil prove the theoretical considerations to be true for MDA-MB-231, MCF-7, T-47-D and ZR-75-1 human breast cancer cell lines in practice.

Key words

Crystal violet chemosensitivity assays Microtitration plates Human breast cancer cell lines Proliferation kinetics Anticancer drug effects 

Abbreviations

PBS

phosphate-buffered saline

FCS

fetal calf serum

NCS

newborn calf serum

ER

oestrogen receptors

PR

progesterone receptor

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Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • Günther Bernhardt
    • 1
  • Herta Reile
    • 1
  • Herbert Birnböck
    • 1
  • Thilo Spruß
    • 1
  • Helmut Schönenberger
    • 1
  1. 1.Institut für PharmazieUniversität RegensburgRegensburgFederal Republic of Germany

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