Abstract
Commercially exploitable compounds are being produced using modern biotechnology for use as food additives, chemotherapeutic agents, and pesticides. Traditionally, animal testing has always played an important role in the safety evaluation of such agents. However, financial and ethical considerations, together with an increased awareness of the limitations of animal models in relation to human metabolism, now warrant the development of alternative testing methods. Therefore, it is fitting that the potential of biotechnology should provide mammalian cell systems for in vitro testing. The ultimate aim of in vitro toxicity testing is the replacement of animals in testing protocols, but in the short term, procedures are refined to reduce the numbers of animals required. This “three Rs” philosophy of reduction, refinement, and replacement was first proposed by Russell and Burch as early as 1959 (1), and is now recognized in the UK Animals in Scientific Procedures Act, 1986 and EC Directive 86/609/ECC (2).
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References
Russell, W. M. S. and Burch, R. L. (1959) The Principles of Humane Experimental Technique, Methuen, London.
Fentem, J. and Balls, M. (1992) In vitro alternatives to toxicity testing in animals. Chem. Ind. 207–211.
Wilson, A. P. (1989) Cytotoxicity and viability assays, in Animal Cell Culture: A Practical Approach (Freshney, R. I., ed.), IRL, Oxford, pp. 183–216.
Freshney, R. I. (1994) Culture of Animal Cells, A Manual of Basic Techniques, Wiley-Liss, New York.
Freshney, R. I. (1989) Animal Cell Culture: A Practical Approach, IRL, Oxford.
Benford, D. J. and Hubbard, S. A. (1987) Preparation and culture of mammalian cells, in Biochemical Toxicology: A Practical Approach (Snell, K. and Mullock, B., eds.), IRL, Oxford, pp. 57–82.
Riddell, R. J., Panacer, D. S., Wilde, S. M., Clothier, R. H., and Balls, M. (1986) The importance of exposure period and cell type in in vitro cytotoxicity testing. ATLA 14, 86–92.
Borenfreund, E. and Puerner, J. A. (1985) Toxicity determined in vitro by morphological alterations and neutral red absorption. Toxicol. Lett. 24, 119–124.
Rotman, B. and Papermaster, B. W. (1966) Membrane properties of living animal cells as studied by enzymatic hydrolysis of fluorogenic esters. Proc. Natl. Acad. Sci. USA 55, 134–141.
Reader, S. J., Blackwell, V., O’Hara, R., Clothier, R. H., Griffin, G., and Balls, M. (1989) A vital dye release method for assaying the short term cytotoxic effects of chemicals and formulations. ATLA 17, 28–33.
Reader, S. J., Blackwell, V., O’Hara, R., Clothier, R. H., Griffin, G., and Balls, M. (1990) Neutral red release from pre-loaded cells as an in vitro approach to testing for eye irritancy potential. Toxicol. In Vitro, 4, 264–266.
Iselt, M, Holtei, W., and Hilgard, P. (1989) The tetrazolium dye assay for rapid in vitro assessment of cytotoxicity. Arzneimittel-forschung 39, 747–749.
Clothier, R. H., Atkinson, K. A., Garle, M. J., Ward, R. K., and Willshaw, A. (1995) The development and evaluation of in vitro tests by the FRAME alternatives laboratory. ATLA 23, 75–90.
Spirlmann, H., Balls, M., Brand, M., Doring, B., Holzhutter, H. G., Kalweit, S., Klecak, G., Eplattenier, H. L., Liebsch, M., Lovell, W. W., Maurer, T., Moldenhauer, F., Moore, L., Pape, W. J. W., Pfanenbecker, U., Potthast, J., Desilva, O., Steiling, W., and Willshaw, A. (1994) EEC COLIPA project on in vitro phototoxicity testing—first results obtained with Balb/c 3T3 cell phototoxicity assay. Toxicol. in Vitro 8, 793–796.
Harbell, J. D., Tsai, Y. C., Maibach, H. I., Gay, R., Miller, K., and Mun, G. C. (1994) An in vivo correlation with 3 in vitro assays to assess skin irritation potential. J. Toxicol. Cut. Ocul. Toxicol. 13, 171–183.
Lewis, M., McCall, J. C., Botham, P. A., and Trebilock, R. (1994) A comparison of 2 cytotoxicity tests for predicting the ocular irritancy of surfactants. Toxicol. in Vitro 8, 867–869.
Christian, M. S. and Diener, R. M. (1996) Soaps and detergents—alternatives to animal eye irritation tests. J. Am. Coll. Toxicol. 15, 1–44.
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© 1999 Humana Press Inc., Totowa, NJ
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Buckberry, L.D. (1999). Cytotoxicity Testing Using Cell Lines. In: Jenkins, N. (eds) Animal Cell Biotechnology. Methods in Biotechnology™, vol 8. Humana Press. https://doi.org/10.1385/0-89603-547-6:239
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DOI: https://doi.org/10.1385/0-89603-547-6:239
Publisher Name: Humana Press
Print ISBN: 978-0-89603-547-8
Online ISBN: 978-1-59259-486-3
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