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A new, rapid colorimetric assay for quantitative determination of cellular proliferation, growth inhibition, and viability

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Journal of tissue culture methods

Summary

A sensitive spectrophotometric test has been developed to rapidly quantitate cell growth and viability on monolayer cultures. The method consists of staining fixed cells with the supravital dye Janus green, extraction of the dye from these cells with absolute alcohol, spectrophotometric measurement of the eluant, and referring optical density (OD) readings to a previously established standard curve such that measurements can be expressed directly as number of surviving cells. Staining of cells with the same colorant before fixation allows microscopical or colorimetric quantitation or both of damaged cells still adherent to the substratum. Both steps can be conveniently carried out on the same cell culture plate. More rapid and easier to perform than the usual cell counting or tritiated thymidine assays, this technique may prove to be useful in evaluation of the effect of growth promoting substances, growth inhibitors, or cytotoxic agents on monolayer cultures.

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V. References

  1. Alcina, A.; Fresno, M. A colorimetric assay based on cell viability for the indirect detection of intracellular replication and killing ofTrypanosoma cruzi. J. Immunol. Methods 105:1–5; 1987.

    Article  CAS  PubMed  Google Scholar 

  2. Artola, A.; Alio, J. L.; Ruiz, J. M., et al. Corneal endothelial cell protection against free radicals by sodium hyaluronate (Healon). Invest. Opthalmol. & Visual Sci. (Suppl.) 32:1177; 1991.

    Google Scholar 

  3. Borenfreund, E.; Puerner, J. A. A simple quantitative procedure using monolayer cultures for cytotoxicity assays (HTD/NR-90). J. Tissue Cult. Methods 9:7–9; 1984.

    Google Scholar 

  4. Gerlier, D.; Thomasset, N. Use of MTT colorimetric assay to measure cell activation. J. Immunol. Methods 94:57–60; 1986.

    Article  CAS  PubMed  Google Scholar 

  5. Givens, K. T.; Kitada, S.; Chen, A. K., et al. Proliferation of human ocular fibroblasts. An assessment of in vitro colorimetric assays. Invest. Ophthalmol. & Visual Sci. 31:1856–1862; 1990.

    CAS  Google Scholar 

  6. Goldmann, R.; Bar-Shavit, Z. Dual effect of normal and stimulated macrophages and their conditioned media on target cell proliferation. JNCI 63:1004–1016; 1979.

    Google Scholar 

  7. Guthoff, R.; Wendl, U.; Böhnke, M., et al. Zur endothelschützenden Wirkung hochvisköser Substanzen in der Kataraktchirurgie. Fortschr. Ophthalmol. In press: 1992.

  8. Hartmann, C.; Rieck, P. A new test for endothelial viability: the Janus green photometry technique. Arch. Ophthalmol. 107:1511–1515; 1989.

    CAS  PubMed  Google Scholar 

  9. Isomura, J.; Yoshimatsu, K.; Ikeda, T., et al. A dye uptake method using cultured human cancer cells and its application to a sensitivity test for anti-cancer drugs. Yakagaku Zasshi. 101:227–231; 1981.

    CAS  Google Scholar 

  10. Landegren, U. Measurement of cell numbers by means of the endogenous enzyme hexosaminidase: applications to detection of lymphokines and cell surface antigens. J. Immunol. Methods 67:379–383; 1984.

    Article  CAS  PubMed  Google Scholar 

  11. Oliver, M. H.; Harrison, N. K.; Bishop, J. E., et al. A rapid and convenient assay for counting cells cultured in microwell plates: application for assessment of growth factors. J. Cell. Sci. 92:513–518; 1989.

    PubMed  Google Scholar 

  12. Mosmann, T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods 67:55–59; 1983.

    Google Scholar 

  13. Nuyts, RMMA.; Edelhauser, H. F.; Pels, E., et al. Toxic effects of detergents on the corneal endothelium. Arch. Ophthalmol. 108:1158–1162; 1990.

    CAS  PubMed  Google Scholar 

  14. Pharmakakis, N.; Hartmann, C.; Bergmann L. In vitro Hornhautendothelschädigung durch intraokulare Linsen aus PMMA, Silikon und Poly-Hema. Fortschr. Ophthalmol. 86:112–115; 1989.

    Google Scholar 

  15. Vaickus, L.; Levy, R. Antiproliferative monoclonal antibodies: detection and initial characterization. J. Immunol. 135:1987–1994; 1985.

    CAS  PubMed  Google Scholar 

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Rieck, P., Peters, D., Hartmann, C. et al. A new, rapid colorimetric assay for quantitative determination of cellular proliferation, growth inhibition, and viability. Journal of Tissue Culture Methods 15, 37–41 (1993). https://doi.org/10.1007/BF02387288

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