3D spheroid culture models for chondrocytes using polyethylene glycol-coated microfabricated chip
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- ARIYOSHI Wataru
- Division of Infections and Molecular Biology, Department of Health Promotion, Kyushu Dental University
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- USUI Michihiko
- Division of Periodontology, Department of Oral Function, Kyushu Dental University
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- SANO Kotaro
- Division of Periodontology, Department of Oral Function, Kyushu Dental University
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- KAWANO Aki
- Division of Infections and Molecular Biology, Department of Health Promotion, Kyushu Dental University
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- OKINAGA Toshinori
- Department of Bacteriology, Osaka Dental University
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- NAKASHIMA Keisuke
- Division of Periodontology, Department of Oral Function, Kyushu Dental University
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- NAKAZAWA Kohji
- Department of Life and Environment Engineering, The University of Kitakyushu
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- NISHIHARA Tatsuji
- Division of Infections and Molecular Biology, Department of Health Promotion, Kyushu Dental University
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抄録
<p>As chondrocytes fail to retain their chondrogenic potential in two-dimensional monolayer cultures, several three-dimensional culture systems have been employed for investigating the physiology and pathophysiology in articular cartilage tissues. In this study, we introduced a polyethylene glycol-coated microfabricated chip that enables spheroid formation from ATDC5 cell line, commonly used as a model for in vitro chondrocyte research. ATDC5 cells cultured in our devices aggregated immediately and generated a single spheroid per well within 24 h. Most cells in spheroids cultured in differentiation medium were viable and the circular shape and smooth surface of the spheroid were maintained up to 14 d in culture. We also detected potent hypoxia conditions, a key factor in chondrogenesis, in whole lesions of ATDC5 spheroids. Expression of chondrogenesis-related genes and type X collagen protein was significantly increased in ATDC5 spheroids grown in differentiation medium, compared with monolayer-cultured ATDC5 cells. We also demonstrated that the differentiation medium-induced Akt protein phosphorylation was upregulated in ATDC5 cells cultured in our spheroid device, suggesting that enhancement of chondrogenic potential in ATDC5 spheroids results from PI3/Akt signaling activation. These results indicated that our spheroid culture system could constitute a high-throughput strategy approach towards elucidating the molecular mechanisms that regulate chondrogenesis.</p>
収録刊行物
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- Biomedical Research
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Biomedical Research 41 (4), 187-197, 2020-08-01
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