RRC ID |
44033
|
著者 |
Newton PT, Staines KA, Spevak L, Boskey AL, Teixeira CC, Macrae VE, Canfield AE, Farquharson C.
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タイトル |
Chondrogenic ATDC5 cells: an optimised model for rapid and physiological matrix mineralisation.
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ジャーナル |
Int J Mol Med
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Abstract |
The development of chondrogenic cell lines has led to major advances in the understanding of how chondrocyte differentiation is regulated, and has uncovered many signalling pathways and gene regulatory mechanisms required to maintain normal function. ATDC5 cells are a well established in vitro model of endochondral ossification; however, current methods are limited for mineralisation studies. In this study we demonstrate that culturing cells in the presence of ascorbic acid and 10 mM β-glycerophosphate (βGP) significantly increases the rate of extracellular matrix (ECM) synthesis and reduces the time required for mineral deposition to occur to 15 days of culture. Furthermore, the specific expression patterns of Col2a1 and Col10a1 are indicative of ATDC5 chondrogenic differentiation. Fourier transform-infrared spectroscopy analysis and transmission electron microscopy (TEM) showed that the mineral formed by ATDC5 cultures is similar to physiological hydroxyapatite. Additionally, we demonstrated that in cultures with βGP, the presence of alkaline phosphatase (ALP) is required for this mineralisation to occur, further indicating that chondrogenic differentiation is required for ECM mineralisation. Together, these results demonstrate that when cultured in the presence of ascorbic acid and 10 mM βGP, ATDC5 cells undergo chondrogenic differentiation and produce a physiological mineralised ECM from Day 15 of culture onwards. The rapid and novel method for ATDC5 culture described in this study is a major improvement compared with currently published methods and this will prove vital in the pursuit of underpinning the molecular mechanisms responsible for poor linear bone growth observed in a number of chronic diseases such as cystic fibrosis, chronic kidney disease, rheumatological conditions and inflammatory bowel disease.
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巻・号 |
30(5)
|
ページ |
1187-93
|
公開日 |
2012-11-1
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DOI |
10.3892/ijmm.2012.1114
|
PMID |
22941229
|
PMC |
PMC3573767
|
MeSH |
Alkaline Phosphatase / antagonists & inhibitors
Alkaline Phosphatase / metabolism
Animals
Calcification, Physiologic*
Cell Differentiation
Cell Line
Chondrocytes / metabolism
Chondrocytes / physiology
Chondrogenesis*
Collagen Type II / genetics
Collagen Type II / metabolism
Collagen Type X / genetics
Collagen Type X / metabolism
Extracellular Matrix / metabolism*
Glycosaminoglycans / metabolism
Levamisole / pharmacology
Mice
Spectroscopy, Fourier Transform Infrared
Transcription, Genetic
|
IF |
3.098
|
引用数 |
42
|
WOS 分野
|
MEDICINE, RESEARCH & EXPERIMENTAL
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リソース情報 |
ヒト・動物細胞 |
|