Université Côte d'Azur, CNRS, UMR7370, LP2M, Nice, France.
Department of Orthopedic Surgery, Baylor College of Medicine, Houston, TX, USA.
Methods Mol Biol. 2021;2308:21-34. doi: 10.1007/978-1-0716-1425-9_2.
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.
骨骼生理学受多种因素调控,包括破骨细胞(OCLs)作为骨吸收细胞、成骨细胞(能够形成骨)、骨细胞或间充质干细胞等重要角色。所有这些细胞彼此之间都有紧密的联系,并影响着不断发生的骨重建过程,以满足骨骼系统不断变化的需求。为了理解这些相互作用,人们必须研究各种细胞类型的孤立功能。然而,OCL 研究存在一个特殊的缺点:由于其巨大的体积、低丰度以及与骨表面的紧密附着,体外分离足够数量的成熟 OCLs 在包括小鼠在内的大多数物种中受到限制或不可行。此外,OCL 可以在体内和体外从不同的祖细胞中获得。因此,体外分化 OCLs 从各种祖细胞仍然是分析 OCL 生物学的关键,这凸显了应用于整个 OCL 研究的可靠金标准方案的重要性。本章将介绍从鼠骨髓细胞、分离的单核细胞和树突状细胞中体外分化 OCLs 的方法,这些方法已经在许多研究中得到验证。