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Alterations in characteristics of canine articular chondrocytes in non-passaged long-term monolayer culture: Matter of differentiation, dedifferentiation and redifferentiation

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Title: Alterations in characteristics of canine articular chondrocytes in non-passaged long-term monolayer culture: Matter of differentiation, dedifferentiation and redifferentiation
Authors: Akaraphutiporn, Ekkapol Browse this author
Sunaga, Takafumi Browse this author →KAKEN DB
Bwalya, Eugene C. Browse this author
Echigo, Ryosuke Browse this author
Okumura, Masahiro Browse this author →KAKEN DB
Keywords: articular chondrocyte
chondrocyte redifferentiation
long-term culture
monolayer culture
tissue engineering
Issue Date: Jun-2020
Publisher: 公益社団法人 日本獣医学会 (The Japanese Society of Veterinary Science)
Journal Title: Journal of veterinary medical science
Volume: 82
Issue: 6
Start Page: 793
End Page: 803
Publisher DOI: 10.1292/jvms.20-0118
Abstract: This study investigated the effects of culture time on phenotype stability of canine articular chondrocytes (CACs) in non-passaged long-term monolayer culture.Third passage (P3) CACs isolated from four cartilage samples were seeded at three different initial seeding densities (0.2 x 10(4),1.0 x 10(4) and 5.0 x 10(4) cells/cm(2)) and maintained in monolayer condition up to 8 weeks without undergoing subculture after confluence. The characteristic changes of chondrocytes during the culture period were evaluated based on the cell morphology, cell proliferation, glycosaminoglycans (GAGs) content, DNA quantification, mRNA expression and ultrastructure of chondrocytes. Chondrocytes maintained under post-confluence condition exhibited a capability to grow and proliferate up to 4 weeks. Alcian blue staining and Dimethylmethylene blue (DMMB) assay revealed that the extracellular matrix (ECM) synthesis was increased in a time-dependent manner from 2 to 8 weeks. The chondrocyte mRNA expression profile was dramatically affected by prolonged culture time, with a significant downregulation of collagen type 1, whereas the expression of collagen type II, aggrecan, Sox9 and matrix metalloproteinase 13 (MMP-13) were significantly upregulated. In addition, transmission electron microscopy (TEM) result indicated dilation of rough endoplasmic reticulum (RER) in these long-term monolayer cultured chondrocytes. These findings demonstrate that the chondrocytes phenotype could be partially redifferentiated through the spontaneous redifferentiation process in long-term cultures using standard culture medium without the addition of chondrogenic supplements or tissue-culture scaffolds.
Rights: https://creativecommons.org/licenses/by-nc-nd/4.0/
Type: article
URI: http://hdl.handle.net/2115/79212
Appears in Collections:獣医学院・獣医学研究院 (Graduate School of Veterinary Medicine / Faculty of Veterinary Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

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