In Vitro Investigation of the Roles of the Proinflammatory Cytokines Tumor Necrosis Factor-α and Interleukin-1 in Murine Osteoclastogenesis
In Vitro Investigation of the Roles of the Proinflammatory Cytokines Tumor Necrosis Factor-α and Interleukin-1 in Murine Osteoclastogenesis
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DOI：
10.1007/978-1-4939-0669-7_10
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发表时间：
2014-01-01
期刊：
影响因子：
--
通讯作者：
Feng, Xu
中科院分区：
文献类型：
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作者：
Jules, Joel;Feng, Xu
虽然单核细胞/巨噬细胞集落刺激因子 (M-CSF) 和 NF kappa B 配体受体激活剂 (RANKL) 对于破骨细胞生成是必需且充分的，但许多其他细胞因子，包括两种促炎细胞因子、肿瘤坏死因子-α (TNF-α) 和白介素-1 (IL-1)，可以对破骨细胞生成过程产生深远的影响。然而，TNF-α 和 IL-1 在破骨细胞生成中的确切作用方式仍存在争议。虽然一些小组证明这两种细胞因子可以在体外仅在 M-CSF 存在的情况下促进小鼠破骨细胞生成，但我们和其他人表明 TNF-α-/IL-1 介导的破骨细胞生成需要允许的 RANKL 水平。本章介绍了我们在使用原代小鼠骨髓巨噬细胞 (BMM) 的体外破骨细胞生成测定中研究 TNF-α 和 IL-1 对破骨细胞形成的影响的方法。提供了详细的实验条件并讨论了关键点，以帮助读者使用该方法独立评估 TNF-α 和 IL-1 在体外破骨细胞生成中的作用。此外，该方法可用于进一步阐明这两种细胞因子与 RANKL 或彼此协同作用以调节破骨细胞生成的信号传导机制。
Whereas the monocyte/macrophage-colony stimulating factor (M-CSF) and the receptor activator of NF kappa B ligand (RANKL) are essential and sufficient for osteoclastogenesis, a number of other cytokines including two proinflammatory cytokines, tumor necrosis factor-alpha (TNF-alpha), and interleukin-1 (IL-1), can exert profound effects on the osteoclastogenic process. However, the precise mode of action of TNF-alpha and IL-1 in osteoclastogenesis remains controversial. While some groups demonstrated that these two cytokines can promote murine osteoclastogenesis in vitro in the presence of M-CSF only, we and others showed that TNF-alpha-/IL-1-mediated osteoclastogenesis requires permissive levels of RANKL. This chapter describes the method that we have used to investigate the effects of TNF-alpha and IL-1 on osteoclast formation in in vitro osteoclastogenesis assays using primary murine bone marrow macrophages (BMMs). Detailed experimental conditions are provided and critical points are discussed to help the reader use the method to independently evaluate the roles of TNF-alpha and IL-1 in osteoclastogenesis in vitro. Moreover, this method can be used to further elucidate the signaling mechanisms by which these two cytokines act in concert with RANKL or with each other to modulate osteoclastogenesis.