Colorimetric growth assay for epidermal cell cultures by their crystal violet binding capacity.

Archives of Dermatological Research
B BonnekohG Mahrle

Abstract

The application of a simple, rapid, and inexpensive colorimetric growth assay was tested for human epidermal cells subcultured in uncoated plastic dishes. Cell layers were incubated with a crystal violet (CV) solution (0.2% with ethanol 2% in 0.5 M Tris-Cl buffer, pH 7.8) for 10 min at room temperature. After rinsing with 0.5 M Tris-Cl (pH 7.8) the cell layer was dried and decolorized with a sodium-dodecylsulfate solution (0.5% with ethanol 50% in 0.5 M Tris-Cl, pH 7.8) for 60 min at 37 degrees C. The extinction of the supernatant was read at the absorption maximum of 586 nm. The protein content of attached cells as classical parameter for quantifying cell growth was strongly related to CV extinction with a correlation coefficient of r = 0.98. Furthermore, the subcellular protein binding qualities of CV were analyzed. The water-soluble protein fraction of cultured epidermal cells was separated by sodium-dodecylsulfate polyacrylamide gel electrophoresis and stained with CV. We found a staining pattern which was qualitatively very similar to that of Coomassie blue, however less intense. Keratin electrophoresis revealed an affinity of CV to the 48, 50, and 56 kD cytokeratins. In conclusion, this CV assay is a reliable and simple m...Continue Reading

References

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Citations

Jan 1, 1991·Archives of Dermatological Research·B FarkasG Mahrle
Nov 26, 2010·Pharmaceutical Research·Michael VosselerRoland Zengerle
Sep 19, 2000·Antimicrobial Agents and Chemotherapy·T V AchenbachR Müller
Dec 14, 2011·Journal of Visualized Experiments : JoVE·Elizabeth J AbrahamSusan Qian
Apr 17, 2009·Journal of Cell Communication and Signaling·Wanhua LuAlexandra E Irvine
Sep 1, 1991·Journal of the American Academy of Dermatology·B BonnekohG Mahrle
Apr 19, 2008·Journal of Neurochemistry·Anayansi Molina-Hernández, Iván Velasco
Apr 9, 2014·The Biochemical Journal·Zheng-Wei LeeLih-Wen Deng
Mar 5, 2015·Biochemical and Biophysical Research Communications·Afag AslanovaMasakazu Yamamoto
Sep 8, 2016·Biochimica Et Biophysica Acta·Camille EttelaieAnthony Maraveyas
Oct 28, 2019·Apoptosis : an International Journal on Programmed Cell Death·Ali M EthaebCamille Ettelaie
Aug 29, 2017·Homeopathy : the Journal of the Faculty of Homeopathy·Jenifer Pendiuk GonçalvesCarolina Camargo de Oliveira
Aug 1, 2019·International Journal of Oncology·Jenifer Pendiuk GonçalvesCarolina Camargo De Oliveira
Nov 25, 2020·Food Chemistry·Fernando Antônio AnjoPaula Toshimi Matumoto-Pintro
Jan 1, 1995·The Journal of Investigative Dermatology·B BonnekohG Mahrle
Sep 16, 2019·Carbohydrate Polymers·Ruberney S OliveiraElaine R Carbonero

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