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Primary mouse osteoblast and osteoclast culturing and analysis.

STAR protocols (2021-04-30)
Claire Chevalier, Melis Çolakoğlu, Julia Brun, Cyril Thouverey, Nicolas Bonnet, Serge Ferrari, Mirko Trajkovski
RÉSUMÉ

Mesenchymal-derived osteoblasts play a key role in bone formation via synthesis and mineralization of the bone and bone remodeling. Osteoclasts are multinucleated cells of hematopoietic origin with a role in bone resorption. Here, we describe a protocol for generating primary cultures of these two cell types from bone tissue including the femur, tibia, and humerus of young mice. We describe methods for addressing their activity and/or differentiation, enabling studying the effects of various treatments during or following differentiation ex vivo. For further practical example of using these protocols, please refer to Chevalier et al. (2020).

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Sigma-Aldrich
4-Nitrophénylphosphate, Phosphate de 4-nitrophényle disodium salt hexahydrate, suitable for enzyme immunoassay, ≥99.0% (enzymatic)
Sigma-Aldrich
2-Phospho-L-ascorbic acid trisodium salt, ≥95.0% (HPLC)
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Chlorure de sodium, BioUltra, for molecular biology, ≥99.5% (AT)
Sigma-Aldrich
Alizarin Red S, certified by the Biological Stain Commission
Sigma-Aldrich
Phosphate de naphthol AS-TR disodium salt, ≥99% (HPLC), Bulk package