Tevlin Ruth, McArdle Adrian, Chan Charles K F, Pluvinage John, Walmsley Graham G, Wearda Taylor, Marecic Owen, Hu Michael S, Paik Kevin J, Senarath-Yapa Kshemendra, Atashroo David A, Zielins Elizabeth R, Wan Derrick C, Weissman Irving L, Longaker Michael T
Hagey Laboratory for Pediatric Regenerative Medicine, Division of Plastic and Reconstructive Surgery, Department of Surgery, Stanford University School of Medicine.
Hagey Laboratory for Pediatric Regenerative Medicine, Division of Plastic and Reconstructive Surgery, Department of Surgery, Stanford University School of Medicine; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University.
J Vis Exp. 2014 Nov 6(93):e52056. doi: 10.3791/52056.
Osteoclasts are highly specialized cells that are derived from the monocyte/macrophage lineage of the bone marrow. Their unique ability to resorb both the organic and inorganic matrices of bone means that they play a key role in regulating skeletal remodeling. Together, osteoblasts and osteoclasts are responsible for the dynamic coupling process that involves both bone resorption and bone formation acting together to maintain the normal skeleton during health and disease. As the principal bone-resorbing cell in the body, changes in osteoclast differentiation or function can result in profound effects in the body. Diseases associated with altered osteoclast function can range in severity from lethal neonatal disease due to failure to form a marrow space for hematopoiesis, to more commonly observed pathologies such as osteoporosis, in which excessive osteoclastic bone resorption predisposes to fracture formation. An ability to isolate osteoclasts in high numbers in vitro has allowed for significant advances in the understanding of the bone remodeling cycle and has paved the way for the discovery of novel therapeutic strategies that combat these diseases. Here, we describe a protocol to isolate and cultivate osteoclasts from mouse bone marrow that will yield large numbers of osteoclasts.
破骨细胞是高度特化的细胞,起源于骨髓中的单核细胞/巨噬细胞谱系。它们独特的吸收骨有机和无机基质的能力意味着它们在调节骨骼重塑中起关键作用。成骨细胞和破骨细胞共同负责动态耦合过程,该过程涉及骨吸收和骨形成共同作用,以在健康和疾病状态下维持正常骨骼。作为体内主要的骨吸收细胞,破骨细胞分化或功能的变化会对身体产生深远影响。与破骨细胞功能改变相关的疾病严重程度不一,从因未能形成造血骨髓空间导致的致命新生儿疾病,到更常见的病症如骨质疏松症,其中过度的破骨细胞骨吸收易导致骨折形成。在体外大量分离破骨细胞的能力使得在理解骨重塑周期方面取得了重大进展,并为发现对抗这些疾病的新治疗策略铺平了道路。在这里,我们描述了一种从小鼠骨髓中分离和培养破骨细胞的方案,该方案将产生大量破骨细胞。