In vitro Cell Viability by CellProfiler ® Software as Equivalent to MTT Assay

• Published in: Pharmacognosy Magazine Vol. 13; no. Suppl 2; pp. S365 - S369
• Main 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
• Format: Journal Article
• Language: English
• Published: India Sage Publications Ltd 01.07.2017; Medknow Publications & Media Pvt Ltd
• Subjects: Algae; Automation; Breast cancer; Cytotoxicity; Fibroblasts; Identification; Liver cancer; Methods; Morphology; Original; Pharmaceutical sciences; Software; Studies; cell viability; CellProfiler; MTT; image analysis; Hepa-1c1c7; L929
• ISSN: 0973-1296; 0976-4062
• DOI: 10.4103/0973-1296.210176

This study evaluated 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. Hepatoma (Hepa-1c1c7) and fibroblast (L929) cells were exposed to isolated substances, camptothecin, lycorine, tazettine, albomaculine, 3-epimacronine, trispheridine, galanthine and , sp. methanolic extract, and 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 . No cytotoxicity at the highest concentration analyzed for 3-epimacronine, albomaculine, galanthine, trispheridine, extract and sp. extract where detected. Tazettine offered cytotoxicity only against the Hepa1c1c7 cell line. Lycorine, camptothecin, and 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 R . 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. cell viability assessment with MTT (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) assay may be replaced by image analysis by CellProfiler . 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 correlation between the two methods was demonstrated by the linear regression plotted as R2. HPLC: High pressure liquid chromatography MTT: (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide).