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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
ABSTRACT

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).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sodium chloride, BioUltra, for molecular biology, ≥99.5% (AT)
Sigma-Aldrich
2-Phospho-L-ascorbic acid trisodium salt, ≥95.0% (HPLC)
Sigma-Aldrich
Naphthol AS-TR phosphate disodium salt, ≥99% (HPLC), Bulk package
Sigma-Aldrich
4-Nitrophenyl phosphate disodium salt hexahydrate, suitable for enzyme immunoassay, ≥99.0% (enzymatic)
Sigma-Aldrich
Alizarin Red S, certified by the Biological Stain Commission