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Culture Expansion in Low-Glucose Conditions Preserves Chondrocyte Differentiation and Enhances Their Subsequent Capacity to Form Cartilage Tissue in Three-Dimensional Culture

Heywood, Hannah K., Nalesso, G, Lee, David A. and Dell'Accio, Francesco (2014) Culture Expansion in Low-Glucose Conditions Preserves Chondrocyte Differentiation and Enhances Their Subsequent Capacity to Form Cartilage Tissue in Three-Dimensional Culture BioResearch Open Access, 3 (1). pp. 9-18.

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Culture 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.

Item Type: Article
Subjects : Veterinary Medicine
Divisions : Faculty of Health and Medical Sciences > School of Veterinary Medicine
Authors :
Heywood, Hannah K.
Nalesso, G
Lee, David A.
Dell'Accio, Francesco
Date : February 2014
DOI : 10.1089/biores.2013.0051
Copyright Disclaimer : © 2012 Mary Ann Liebert, Inc. publishers. All rights reserved, USA and worldwide. BioResearch Open Access publishes articles under the liberal CC-BY license. This means that articles can be freely redistributed and reused by the author and others as long as the article is properly cited. Published articles in BioResearch Open Access can be deposited immediately into an online repository or social network without an embargo. BioResearch Open Access articles can be emailed to colleagues, printed, archived in a collection, included in course-packs, and distributed without restrictions. Please read the full Creative Commons license for further information.
Uncontrolled Keywords : Chondrocyte, Crabtree effect, Differentiation markers, Glucose concentration, Monolayer expansion
Depositing User : Symplectic Elements
Date Deposited : 07 Apr 2017 11:45
Last Modified : 31 Oct 2017 19:18

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