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- Title
In vitro Cell Viability by CellProfiler® Software as Equivalent to MTT Assay.
- Authors
Gasparini, Luciana S.; Macedo, Nayana D.; Pimentel, Elisângela F.; Fronza, Marcio; Junior, Valdemar L.; Borges, Warley S.; Cole, Eduardo R.; Andrade, Tadeu U.; Endringer, Denise C.; Lenz, Dominik
- Abstract
Objective: This study evaluated in vitro cell viability by the colorimetric MTT stands for 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) assay compared to image analysis by CellProfiler® software. Materials and Methods: Hepatoma (Hepa-1c1c7) and fibroblast (L929) cells were exposed to isolated substances, camptothecin, lycorine, tazettine, albomaculine, 3-epimacronine, trispheridine, galanthine and Padina gymnospora, Sargassum sp. methanolic extract, and Habranthus itaobinus Ravenna ethyl acetate in different concentrations. After MTT assay, cells were stained with Panotic dye kit. Cell images were obtained with an inverted microscope equipped with a digital camera. The images were analyzed by CellProfiler®. Results: No cytotoxicity at the highest concentration analyzed for 3-epimacronine, albomaculine, galanthine, trispheridine, P. gymnospora extract and Sargassum sp. extract where detected. Tazettine offered cytotoxicity only against the Hepa1c1c7 cell line. Lycorine, camptothecin, and H. itaobinus extract exhibited cytotoxic effects in both cell lines. The viability methods tested were correlated demonstrated by Bland-Atman test with normal distribution with mean difference between the two methods close to zero, bias value 3.0263. The error was within the limits of the confidence intervals and these values had a narrow difference. The correlation between the two methods was demonstrated by the linear regression plotted as R². Conclusion: CellProfiler® image analysis presented similar results to the MTT assay in the identification of viable cells, and image analysis may assist part of biological analysis procedures. The presented methodology is inexpensive and reproducible.
- Subjects
CELL survival; COLORIMETRIC analysis; IMAGE analysis; HEPATOCELLULAR carcinoma; CAMPTOTHECIN; FIBROBLASTS
- Publication
Pharmacognosy Magazine, 2017, Vol 13, pS365
- ISSN
0973-1296
- Publication type
Article
- DOI
10.4103/0973-1296.210176