Chondrogenic ATDC5 cells: An optimised model for rapid and physiological matrix mineralisation

Newton,P. T.; Staines,K. A.; Spevak,L.; Boskey,A.  L.; Teixeira,C. C.; Macrae,V. E.; Canfield,A.  E.; Farquharson,C.

doi:10.3892/ijmm.2012.1114

November 2012 Volume 30 Issue 5

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November 2012 Volume 30 Issue 5

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Article

Chondrogenic ATDC5 cells: An optimised model for rapid and physiological matrix mineralisation

Authors:
- P. T. Newton
- K. A. Staines
- L. Spevak
- A. L. Boskey
- C. C. Teixeira
- V. E. Macrae
- A. E. Canfield
- C. Farquharson
View Affiliations / Copyright

Affiliations: The Roslin Institute and R(D)SVS, University of Edinburgh, Easter Bush, Midlothian EH25 9RG, UK, Musculoskeletal Integrity Program, Hospital for Special Surgery, New York, NY, USA, Basic Science and Craniofacial Biology, New York University, College of Dentistry, New York, NY, USA, Wellcome Trust Centre for Cell-Matrix Research and Cardiovascular Research Group, Manchester M13 9PT, UK
Pages: 1187-1193
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Published online on: August 31, 2012

https://doi.org/10.3892/ijmm.2012.1114
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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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Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Chondrogenic ATDC5 cells: An optimised model for rapid and physiological matrix mineralisation

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