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3D spheroid culture models for chondrocytes using polyethylene glycol-coated microfabricated chip.
- Source :
-
Biomedical research (Tokyo, Japan) [Biomed Res] 2020; Vol. 41 (4), pp. 187-197. - Publication Year :
- 2020
-
Abstract
- 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.
- Subjects :
- Animals
Biomarkers metabolism
Cell Differentiation drug effects
Cell Hypoxia genetics
Cell Line
Chondrocytes cytology
Chondrocytes metabolism
Chondrogenesis drug effects
Chondrogenesis genetics
Coated Materials, Biocompatible chemistry
Collagen Type II genetics
Collagen Type II metabolism
Collagen Type X genetics
Collagen Type X metabolism
Gene Expression
Hyaluronan Synthases genetics
Hyaluronan Synthases metabolism
Lab-On-A-Chip Devices
Mice
Platinum chemistry
Polymethyl Methacrylate chemistry
Spheroids, Cellular cytology
Spheroids, Cellular metabolism
Cell Culture Techniques
Cell Proliferation drug effects
Chondrocytes drug effects
Coated Materials, Biocompatible pharmacology
Polyethylene Glycols pharmacology
Spheroids, Cellular drug effects
Subjects
Details
- Language :
- English
- ISSN :
- 1880-313X
- Volume :
- 41
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Biomedical research (Tokyo, Japan)
- Publication Type :
- Academic Journal
- Accession number :
- 32801268
- Full Text :
- https://doi.org/10.2220/biomedres.41.187