Abstract
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.
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References
AH Cory TC Owen JA Barltrop JG Cory (1991) ArticleTitleUse of an aqueous soluble tetrazolium/formazan assay for cell growth assay in culture Cancer Commun. 3 207–212
S Guerra ParticleDel C Bracci K Nilson et al. (2001) ArticleTitleEntrapment of dispersed pancreatic islet cells in Cultispher-S macroporous gelatin microcarriers: preparation, in vitro characterization, and microencapsulation Biotechnol. Bioeng. 75 741–744
C Frondoza A Sohrabi D Hungerford (1996) ArticleTitleHuman chondrocytes proliferate and produce matrix components in microcarrier suspension culture Biomaterials 17 879–888
O Gurevich A Vexler G Marx T Prigozhina L Levdansky S Slavin I Shimeliovich R Gorodetsky (2002) ArticleTitleFibrin microbeads for isolatin and growing bone marrow-derived progenitor cells capable of forming bone tissue Tissue Eng. 8 661–672
K Lappalainen I Jaaskelainen K Syrjanen A Urtti S Syrjanen (1994) ArticleTitleComparison of cell proliferation and toxicity using two cationic liposomes Pharm. Res. 11 1127–1131
J Malda CA Blitterswijk Particlevan M Grojec DE Martens J Tramper J Riesle (2003) ArticleTitleExpansion of bovine chondrocytes on microcarriers enhances redifferentiation Tissue Eng. 9 939–948
K McConnell M Jarman-Smith K Stewart JB Chaudhuri (2004) ArticleTitleCulture of meniscal chondrocytes on alginate hydrogel matrices Food Bioprod. Process. 82 126–132
T Mosmann (1983) ArticleTitleRapid colorimetric assay for cellular growth and survival; application to proliferation and cytotoxicity assay J. Immunol. Methods 65 55–63
JW Muhitch KC O’Connor DA Blake DJ Lacks N Rosenzweig GF Spaulding (2000) ArticleTitleCharacterization of aggregation and protein expression of bovine corneal endothelial cells as microcarrier cultures in a rotating wall vessel Cytotechnology 32 253–263
KK Sanford WR Earle VJ Evans HK Waitz JE Shannon (1951) ArticleTitleThe measurement of proliferation in tissue cultures by enumeration of cell nuclei J. Natl. Cancer Inst. 11 773–795
JH Taylor PS Woods WL Hughes (1957) ArticleTitleThe organization and duplication of chromosomes as revealed by autoradioagraphic studies using tritium labeled thymidine Proc. Natl. Acad. Sci. USA 43 122–128
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Pabbruwe, M.B., Stewart, K. & Chaudhuri, J.B. A Comparison of Colorimetric and DNA Quantification Assays for the Assessment of Meniscal Fibrochondrocyte Proliferation in Microcarrier Culture. Biotechnol Lett 27, 1451–1455 (2005). https://doi.org/10.1007/s10529-005-1308-x
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DOI: https://doi.org/10.1007/s10529-005-1308-x