Differentiation and Phenotyping of Murine Osteoclasts from Bone Marrow Progenitors, Monocytes, and Dendritic Cells

Bone physiology is dictated by various players, including osteoclasts (OCLs) as bone resorbing cells, osteoblasts (capable of bone formation), osteocytes, or mesenchymal stem cells, to mention the most important players. All these cells are in tight communication with each other and influence the constantly occurring process of bone remodeling to meet changing requirements on the skeletal system. In order to understand these interplays, one must investigate isolated functions of the various cell types. However, OCL research displays a special drawback: due to their giant size, low abundance, and tight attachment on the bone surface, ex vivo isolation of sufficient amounts of mature OCLs is limited or not conceivable in most species including mice. Moreover, OCLs can be obtained from different progenitors in vivo as well as in vitro. Thus, in vitro differentiation of OCLs from various progenitor cells remains essential in the analysis of OCL biology, underlining the importance of reliable gold standard protocols to be applied throughout OCL research. This chapter will deal with in vitro differentiation of OCLs from murine bone marrow cells, as well as isolated monocytes and dendritic cells that have already been validated in numerous studies.