3D spheroid culture models for chondrocytes using polyethylene glycol-coated microfabricated chip

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-coat...

Full description

Saved in:
Bibliographic Details
Published in:Biomedical Research 2020/08/01, Vol.41(4), pp.187-197
Main Authors: ARIYOSHI, Wataru, USUI, Michihiko, SANO, Kotaro, KAWANO, Aki, OKINAGA, Toshinori, NAKASHIMA, Keisuke, NAKAZAWA, Kohji, NISHIHARA, Tatsuji
Format: Article
Language:English
Subjects:
AKT protein
Cartilage
Cartilage (articular)
Cell culture
Cell differentiation
Chondrocytes
Chondrogenesis
Collagen
Differentiation
Gene expression
Hypoxia
Molecular modelling
Monolayers
Phosphorylation
Polyethylene glycol
Proteins
Spheroids
Three dimensional models
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:0388-6107
1880-313X
DOI:10.2220/biomedres.41.187