基础医不学与临床该稿1.docVIP

题名： 细胞因子诱导破骨细胞分化的两种实验模型的建立 姓名：蒋东芳 工作单位：北京人民医院关节病诊疗中心 通讯作者：蒋东芳 通讯地址：100044 北京西直门南大街11号 人民医院骨关节实验室 电话 传真号Email：drdrj@ 细胞因子诱导破骨细胞分化的两种实验模型的建立 蒋东芳, 吕厚山, 林剑浩, 姜军, 陈占昆 （北京大学 人民医院 关节病诊疗中心， 北京 100044） 摘要： 目的 探讨体外从单核细胞诱导分化形成破骨细胞的两种不 同方法，建立研究细胞因子对破骨细胞分化作用的两种不同实验模 型。方法 外周血单核细胞在10-8mol/l LTB4和25μg/L M-CSF刺 激下，经过2周的直接诱导分化；外周血单核细胞与RAFLs共培养， 加入10-8 mol/l LTB4和25μg/L M-CSF，经过3周的间接诱导分化， 用抗酒石酸酸性磷酸酶(TRAP)染色鉴定破骨细胞样细胞(OCL)。结果 通过直接分化和间接分化均可形成破骨细胞样细胞(OCL)。结论 两 种不同实验模型能分别用于研究细胞因子对破骨细胞的直接和间接 分化作用。 关键词： 单核细胞；破骨细胞；细胞因子；实验模型 中号:图分类号：Q2 文献标识码：A Establishment of two experimental models for studying the osteoclast differentiation induced by Cytokines JIANG Dong-fang, LU Hou-shan, LIN Jian-hao, JIANG Jun, CHEN Zhan-kun (Arthritis ClinicResearch Center, Peking University People Hospital, Beijing 100044, China ) Abstract: Objective Establishment of two experimental models for osteoclast differentiation of monocyte from invitro, and study the potential of osteoclast differentiation induced by cytokines. Method CD14+ monocyte fraction of peripheral blood mononuclear cell (PBMC) stimulated by 25ng/ml M-CSF+10-8M LTB4 for twoweeks. Indirectly stimulated by 25μg/L M-CSF+10-8mol/L LTB4 for two weeks. Indirectly model of osteoclast differentiation: Utilize the coculture model of RAFLs and monocyte that were stimulated in the presence of 25μg/L M-CSF+10-8mol/L LTB4 for three weeks. Through TRAP staining we observed result of multinucleated TRAP staining positive osteoclast-like cells as differentiation effect of each group. Results Osteoclast-like cells can be induced by both direct and indirect models. Discuss Two experimental models for osteoclast differentiation can be used to study effect of various cytokines on directly and indirectly OC differentiation. Key word：Monocyte；Osteoclast；Cytokine；experimental model 破骨细胞（Osteoclasts/OC）是人体生理性骨重建和病理性骨破坏 （如RA、牙周疾病、骨转移癌）过程中高度特异性的唯一具有骨质 吸收功能的多核性细胞，来源于造血前体细胞 。RANKL和M-CSF 是破骨细胞形