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大鼠骨髓破骨细胞前体的分离、纯化及分化

Isolation, Purification, and Differentiation of Osteoclast Precursors from Rat Bone Marrow.

作者信息

Wang Lining, Zheng Suyang, Guo Yang, Pan Yalan, Sun Jie, Xu Weimin, Lu Jinlan, Li Weidong, Ma Yong

机构信息

Laboratory of New Techniques of Restoration & Reconstruction, Institute of Traumatology & Orthopedics, Nanjing University of Chinese Medicine; TCM Nursing Intervention Laboratory of Chronic Disease Key Laboratory, Nanjing University of Chinese Medicine.

Jiangsu Key Laboratory of Chinese Medicine Processing, Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing, Nanjing University of Chinese Medicine.

出版信息

J Vis Exp. 2019 May 19(147). doi: 10.3791/58895.

Abstract

Osteoclasts are large, multinucleated, and bone-resorbing cells of the monocyte-macrophage lineage that are formed by the fusion of monocytes or macrophage precursors. Excessive bone resorption is one the most significant cellular mechanisms leading to osteolytic diseases, including osteoporosis, periodontitis, and periprosthetic osteolysis. The main physiological function of osteoclasts is to absorb both the hydroxyapatite mineral component and the organic matrix of bone, generating the characteristic resorption appearance on the surface of bones. There are relatively few osteoclasts compared to other cells in the body, especially in adult bones. Recent studies have focused on how to obtain more mature osteoclasts in less time, which has always been a problem. Several improvements in the isolation and culture techniques have developed in laboratories in order to obtain more mature osteoclasts. Here, we introduce a method that isolates bone marrow in less time and with less effort compared to the traditional procedure, using a special and simple device. With the use of density gradient centrifugation, we obtain large amounts of fully differentiated osteoclasts from rat bone marrow, which are identified by classical methods.

摘要

破骨细胞是单核巨噬细胞谱系中大型的、多核的骨吸收细胞,由单核细胞或巨噬细胞前体融合形成。过度的骨吸收是导致溶骨性疾病(包括骨质疏松症、牙周炎和假体周围骨溶解)的最重要细胞机制之一。破骨细胞的主要生理功能是吸收骨的羟基磷灰石矿物质成分和有机基质,在骨表面产生特征性的吸收外观。与体内其他细胞相比,破骨细胞数量相对较少,尤其是在成年骨骼中。最近的研究集中在如何在更短的时间内获得更多成熟的破骨细胞,这一直是个问题。为了获得更多成熟的破骨细胞,实验室已经开发了几种分离和培养技术的改进方法。在这里,我们介绍一种方法,与传统方法相比,使用一种特殊且简单的装置,可以在更短的时间内、更轻松地分离骨髓。通过使用密度梯度离心法,我们从大鼠骨髓中获得了大量完全分化的破骨细胞,并通过经典方法进行了鉴定。

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