A comparison of colorimetric and DNA quantification assays for the assessment of meniscal fibrochondrocyte proliferation in microcarrier culture.
We have developed and refined a rapid, reliable method for the evaluation of attachment and proliferation of ovine meniscal chondrocytes in microcarrier culture. Assays measuring both mitochondrial activity, using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] and MTS [3-(4,5-dimethylthiazol-2-yl)5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium], and DNA synthesis with a PicoGreen assay were compared. The MTT assay was the most sensitive at lower cell concentrations and enabled accurate assessment of cell proliferation over 14 day culture.
Pabbruwe
Moreica B
MB
Centre for Regenerative Medicine, Department of Chemical Engineering, University of Bath, BA2 7AY, Bath, United Kingdom.
Stewart
Karina
K
Chaudhuri
Julian B
JB
D000818:Animals; D001681:Biological Assay / Q000379:methods; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology*; D003124:Colorimetry / Q000379:methods*; D004247:DNA / Q000096:biosynthesis; D051445:Fibrocartilage / Q000166:cytology*; D008928:Mitochondria / Q000378:metabolism; D012680:Sensitivity and Specificity; D012756:Sheep; D013778:Tetrazolium Salts; D013844:Thiazoles
D013778:Tetrazolium Salts; D013844:Thiazoles; C070380:3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium; D004247:DNA; C022616:thiazolyl blue