Skip to main content
SpringerLink
Log in

A simple quantitative procedure using monolayer cultures for cytotoxicity assays (HTD/NR-90)

  • Published:
Journal of tissue culture methods

Summary

A sensitive quantitative procedure for assaying viable cells in monolayer cultures is described. The two-component test involves (a) microscopic screening for morphological alterations after an experimental protocol for cytotoxicity studies and (b) quantitation of surviving cells by incubation with the supravital dye neutral red, followed by colorimetric analysis of the dye extracted from the lysosomes of the viable cells. Both assays, conveniently carried out within the same culture, have been standardized for use with 96-well microtiter plates and automatic reading with a Dynatech spectrophotometric microplate reader. The test can be adapted for use with conventional spectrophotometers.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Bitensky, L. The reversible activation of lysosomes in normal cells and the effect of pathological conditions. In: Ciba Foundation Symposium “Lysosomes” de Reuck, A. V. S. and Cameron, M. P., eds. Boston, MA: Little, Brown and Co. 1963: 362–375.

    Google Scholar 

  2. Borenfreund, E.; Borrero, O. In vitro cytotoxicity assays: Potential alternatives to the Draize ocular irritancy test. Cell Biol. Toxicol. 1: 55–65; 1984.

    Google Scholar 

  3. Borenfreund, E.; Puerner, J. A. Toxicity determined in vitro by morphological alternation and neutral absorption. Toxicol. Lett. 24:119–124; 1985.

    Google Scholar 

  4. Borenfreund, E.; Shopsis, C. Toxicity monitored with a correlated set of cell culture assays. Xenobiotica 1985 in press.

  5. Lullmann-Rauch, R. Drug induced lysosomal storage disorders. In: Dingle, J. T.; Jacques, P. J.; Shaw, I. H., eds. Lysosomes in applied biology and therapeutics, vol. 6 Amsterdam: North Holland Publ. Co.; 1979: 49–129.

    Google Scholar 

  6. Nemes, Z.; Dietz, R.; Luth, J. B.; et al. The pharmacological relevance of vital staining with neutral red. Experientia 35:1475–1476; 1979.

    Google Scholar 

  7. Winckler, J. Vital staining of lysosomes and other cell organelles of the rat with neutral red. Prog. Histochem. Cytochem. 6:1–89; 1974.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratory Animal Research Center, The Rockefeller University, 10021, New York, N. Y., USA

    Ellen Borenfreund & James A. Puerner

Authors
  1. Ellen Borenfreund
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. James A. Puerner
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Borenfreund, E., Puerner, J.A. A simple quantitative procedure using monolayer cultures for cytotoxicity assays (HTD/NR-90). Journal of Tissue Culture Methods 9, 7–9 (1985). https://doi.org/10.1007/BF01666038

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01666038

Key words

Search

Navigation