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Protocol no. 36
CHINESE HAMSTER OVARY (CHO) CELL PROLIFERATION TEST

The inhibition of CHO cell proliferation provides an overall assessment of the toxicity of the test substance.

CONTACT

Dr. Ingolf Cascorbi
Free University of Berlin Institute of Biochemistry and Molecular Biology Ehrenbergsstr. 26-28 D-W-1000 Berlin 33 Germany

RATIONALE

The rate of cellular proliferation may be regarded as an overall indicator of the physiological status of the cell. Therefore, the effect of various toxic substances on different cell functions will be reflected by changes in the proliferation rate.

BASIC PROCEDURE

CHO cells are cultured for one week in petri dishes with various concentrations of test substance. The number of cells is counted twice daily, and the proliferation rate is determined from the logarithmic growth phase. A dose response curve is obtained by plotting relative growth rates versus concentration of chemical.

CRITICAL ASSESSMENT

CHO cells are a well characterized, sensitive and reliable mammalian test system. Their proliferation rate has been found to correlate with physiological membrane functions, such as adenosine uptake and the activity of Na+/K+-ATPase (Forêt and Ahlers, 1988; Cascorbi and Ahlers, 1989). The fact that significant correlations have been found between toxicity assessed by the proliferation rate of CHO cells and results from aquatic test systems indicates the potential use of this system for ecotoxicological studies (see Ahlers et al, 1991 for a comparison of yeast and CHO cells; other comparisons with data from fish, daphnia and algae have also been carried out). The chief disadvantage associated with this method is the need to wait one week to get the results.

TEST STATUS

The test system has been validated in research studies supported by the German Federal Environmental Agency.

CHEMICALS TESTED

2,4-Dichlorophenol 2,6-Dichlorophenol 3,5-Dimethoxyphenol 4-Methyl-2-nitrophenol 2,3,4,5-tetrachlorophenol 2,4,6-Trichlorophenol 3-(Trifluoromethyl)phenol 2,4,6-Triiodophenol ORGANIZATIONS USING THE SYSTEM In-house use.

REFERENCES

  1. Ahlers, J., Cascorbi, I., Forêt, M., Gies, A., Köhler, M., Pauli, W. & Rösick, E. (1991) Interaction with functional membrane proteins - a common mechanism of toxicity for lipophilic environmental chemicals? Comp. Physiol. Biochem. 100C, 111-113.
  2. Cascorbi, I. (1989) Na+/K+-ATPase of mammalian cells as test system for predicting chemical hazard of environmental chemicals. A QSAR study. Biol. Chem. Hoppe Seyler 370, 617.
  3. Cascorbi, I. & Ahlers, J. (1989) Correlation between the lipophilicity of substituted phenols and their inhibition of the Na+/K+-ATPase of Chinese hamster ovary cells. Toxicology 58, 197-210.
  4. Forêt, M. & Ahlers, J. (1988) Effects of phenols on growth rate and adenosine uptake of CHO cells. Ecotoxicol. Environ. Safety 16, 303-309.

IP-36 © November 1991