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Studies of mineralization in tissue culture: optimal conditions for cartilage calcificationThe optimal conditions for obtaining a calcified cartilage matrix approximating that which exists in situ were established in a differentiating chick limb bud mesenchymal cell culture system. Using cells from stage 21-24 embryos in a micro-mass culture, at an optimal density of 0.5 million cells/20 microliters spot, the deposition of small crystals of hydroxyapatite on a collagenous matrix and matrix vesicles was detected by day 21 using X-ray diffraction, FT-IR microscopy, and electron microscopy. Optimal media, containing 1.1 mM Ca, 4 mM P, 25 micrograms/ml vitamin C, 0.3 mg/ml glutamine, no Hepes buffer, and 10% fetal bovine serum, produced matrix resembling the calcifying cartilage matrix of fetal chick long bones. Interestingly, higher concentrations of fetal bovine serum had an inhibitory effect on calcification. The cartilage phenotype was confirmed based on the cellular expression of cartilage collagen and proteoglycan mRNAs, the presence of type II and type X collagen, and cartilage type proteoglycan at the light microscopic level, and the presence of chondrocytes and matrix vesicles at the EM level. The system is proposed as a model for evaluating the events in cell mediated cartilage calcification.
Document ID
20050000829
Document Type
Reprint (Version printed in journal)
Authors
Boskey, A. L. (Hospital for Special Surgery New York, NY 10021)
Stiner, D.
Doty, S. B.
Binderman, I.
Leboy, P.
Date Acquired
August 22, 2013
Publication Date
January 1, 1992
Publication Information
Publication: Bone and mineral
Volume: 16
Issue: 1
ISSN: 0169-6009
Subject Category
Life Sciences (General)
Funding Number(s)
CONTRACT_GRANT: DE 08239
CONTRACT_GRANT: AR 37661
Distribution Limits
Public
Copyright
Other
Keywords
Non-NASA Center
NASA Discipline Musculoskeletal