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dc.contributor.authorHeywood, HKen_US
dc.contributor.authorNalesso, Gen_US
dc.contributor.authorLee, DAen_US
dc.contributor.authorDell'accio, Fen_US
dc.description.abstractCulture conditions that preserve a stable chondrocyte phenotype are desirable in cell-based cartilage repair to maximize efficacy and clinical outcome. This study investigates whether low-glucose conditions will preserve the chondrocyte phenotype during culture expansion. Articular chondrocytes were culture-expanded in media supplemented with either low (1 mM) or high (10 mM) glucose. The metabolic phenotype, reactive oxygen species generation, and mRNA expression of markers of differentiation or catabolism were assessed by reverse-transcription quantitative polymerase chain reaction after four population doublings (PDs) and subsequent tissue formation capacity determined using pellet cultures. Continuous monolayer culture was used to determine the population doubling limit. After expansion in monolayer for four PDs, chondrocytes expanded in low-glucose conditions exhibited higher expression of the differentiation markers SOX9 and COL2A1 and reduced expression of the catabolic metalloproteinase matrix metallopeptidase 13. When chondrocytes expanded in low glucose were cultured in micropellets, they consistently generated more cartilaginous extracellular matrix than those expanded in high glucose, as evaluated by wet weight, sulfated glycosaminoglycan content, and hydroxyproline assay for collagen content. The same pattern was observed whether high or low glucose was used during the pellet culture. During expansion, chondrocytes in high-glucose generated 50% more reactive oxygen species than low-glucose conditions, despite a lower dependence on oxidative phosphorylation for energy. Furthermore low-glucose cells exhibited >30% increased population doubling limit. These data suggests that low-glucose expansion conditions better preserve the expression of differentiation markers by chondrocytes and enhance their subsequent capacity to form cartilage in vitro. Therefore, low glucose levels should be considered for the expansion of chondrocytes intended for tissue engineering applications.en_US
dc.description.sponsorshipThis study was funded by the Medical Research Council/Engineering and Physical Sciences Research Council (EPSRC) discipline bridging initiative grant PPA026, EPSRC Platform Grant EP/E046975/1; Human Frontier Science Program Grant RGP0025/2009-C and Arthritis Research U.K. grants 19654 and 19344.en_US
dc.format.extent9 - 18en_US
dc.relation.ispartofBiores Open Accessen_US
dc.rightsCC0 1.0 Universal*
dc.subjectCrabtree effecten_US
dc.subjectdifferentiation markersen_US
dc.subjectglucose concentrationen_US
dc.subjectmonolayer expansion potentialen_US
dc.subjectregenerative potentialen_US
dc.subjecttissue engineeringen_US
dc.titleCulture expansion in low-glucose conditions preserves chondrocyte differentiation and enhances their subsequent capacity to form cartilage tissue in three-dimensional culture.en_US
pubs.notesNot knownen_US
qmul.funderTargeting WNT/CaMKII in Osteoarthritis::MRCen_US

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CC0 1.0 Universal
Except where otherwise noted, this item's license is described as CC0 1.0 Universal