TY - JOUR
T1 - Application and evaluation of the alamarblue assay for cell growth and survival of fibroblasts
AU - Voytik-Harbin, Sherry L.
AU - Brightman, A. O.
AU - Waisner, B.
AU - Lamar, C. H.
AU - Badylak, S. F.
N1 - Funding Information:
We thank Dr. J. P. Robinson and his staff (Kathy Ragheb, Steve Kelley, and Gretchen Lawler) for generously sharing their laboratory, space and equipment so that these studies could be conducted. In addition, we thank Debra Klich for her technical support and Dr. John Turek and Dr. Tim McPherson for their comments and suggestions. This work was supported by funding obtained from NIH (HD31425-01) by S. E B. and Purdue University (TRASK FUND) by S. F. B. and S. L. V.-H.
PY - 1998
Y1 - 1998
N2 - 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.
AB - 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.
KW - AlamarBlue
KW - Bioassay
KW - Fibroblast
KW - Growth factor assay
KW - Proliferation
UR - http://www.scopus.com/inward/record.url?scp=0032468181&partnerID=8YFLogxK
U2 - 10.1007/s11626-998-0130-x
DO - 10.1007/s11626-998-0130-x
M3 - Article
AN - SCOPUS:0032468181
SN - 1071-2690
VL - 34
SP - 239
EP - 246
JO - In Vitro Cellular and Developmental Biology-Animal
JF - In Vitro Cellular and Developmental Biology-Animal
IS - 3
ER -