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Application and evaluation of the alamarblue assay for cell growth and survival of fibroblasts

  • Sherry L. Voytik-Harbin
  • Andrew O. Brightman
  • Beverly Waisner
  • Carlton H. Lamar
  • Stephen F. Badylak
Growth, Differentiation And Senescence

Summary

Cell proliferation assays are essential to developing an understanding of the molecular mechanisms that modulate cell growth and differentiation. In this paper, we describe the application of alamarBlue, a new and versatile metabolic dye, for the detection of Swiss 3T3 fibroblast proliferation and/or survival. As a redox indicator, alamarBlue is reduced by reactions innate to cellular metabolism and, therefore, provides an indirect measure of viable cell number. Various assay parameters were optimized for a 96-well format to achieve a detectable range of fibroblast cell number from 100 to 20 000 cells/well, which is similar to that obtained with traditional (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and [3H]thymidine assay techniques. Standard (reference) curves generated with a known fibroblast stimulator were used to facilitate quantitation and comparison of unknown test substances. The alamarBlue assay offers the advantages of technical simplicity, freedom from radioisotopes, versatility in detection, no extraction, and excellent reproducibility and sensitivity. We anticipate that this simple and versatile alamarBlue assay, when used alone or in conjunction with other bioassays, will be a useful tool for investigating the complex mechanisms of cellular proliferation.

Key words

alamarBlue bioassay growth factor assay fibroblast proliferation 

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Copyright information

© Society for In Vitro Biology 1998

Authors and Affiliations

  • Sherry L. Voytik-Harbin
    • 1
  • Andrew O. Brightman
    • 1
  • Beverly Waisner
    • 2
  • Carlton H. Lamar
    • 2
  • Stephen F. Badylak
    • 1
  1. 1.Hillenbrand Biomedical Engineering CenterPurdue UniversityWest Lafayette
  2. 2.Department of Basic Medical Sciences, School of Veterinary MedicinePurdue UniversityWest Lafayette

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