A three-dimensional tissue culture model of bone formation utilizing rotational co-culture of human adult osteoblasts and osteoclasts
A three-dimensional tissue culture model of bone formation utilizing rotational co-culture of human adult osteoblasts and osteoclasts
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DOI：
10.1016/j.actbio.2013.04.051
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发表时间：
2013-08-01
影响因子：
9.7
通讯作者：
Pellis, Neal R.
中科院分区：
文献类型：
--
作者：
Clarke, Mark S. F.;Sundaresan, Alamelu;Pellis, Neal R.
活骨是一种复杂的三维复合材料，由矿化的细胞外基质中空间组织的多种细胞类型组成。迄今为止，复杂的细胞水平的串扰之间的主要骨形成细胞参与骨的机械和生物化学刺激的反应的机制研究一直受到阻碍，缺乏一个合适的体外模型，捕捉这种反应的“耦合”的性质。使用一种新的旋转共培养方法，我们从正常人原代成骨细胞和破骨细胞前体细胞的混合物中产生了大的（直径>4 mm）三维矿化组织构建体，而不需要任何可能干扰这种细胞-细胞相互作用的外源性骨传导支架材料。成熟、分化的骨结构由破骨细胞和成骨细胞居住的外部区域和包含包裹在自组装的多孔矿化细胞外基质中的骨细胞的中心区域组成。骨构建体表现出与重塑人小梁骨相似的形态学、矿物质和生物化学特征，包括构建体内SOST、BGT、ACP 5、BMP-2、BMP-4和BMP-7的mRNA表达以及BMP-2蛋白分泌到培养基中。这种骨形成的“耦合”模型将允许对正常骨形成/重塑过程的各种刺激的未来研究，因为它涉及成骨细胞、破骨细胞和骨细胞在人类矿化组织生成中的细胞功能。(C)2013 Acta Materialia Inc.由爱思唯尔有限公司出版。保留所有权利。
Living bone is a complex, three-dimensional composite material consisting of numerous cell types spatially organized within a mineralized extracellular matrix. To date, mechanistic investigation of the complex cellular level cross-talk between the major bone-forming cells involved in the response of bone to mechanical and biochemical stimuli has been hindered by the lack of a suitable in vitro model that captures the "coupled" nature of this response. Using a novel rotational co-culture approach, we have generated large (>4 mm diameter), three-dimensional mineralized tissue constructs from a mixture of normal human primary osteoblast and osteoclast precursor cells without the need for any exogenous osteoconductive scaffolding material that might interfere with such cell-cell interactions. Mature, differentiated bone constructs consist of an outer region inhabited by osteoclasts and osteoblasts and a central region containing osteocytes encased in a self-assembled, porous mineralized extracellular matrix. Bone constructs exhibit morphological, mineral and biochemical features similar to remodeling human trabecular bone, including the expression of mRNA for SOST, BGLAP, ACP5, BMP-2, BMP-4 and BMP-7 within the construct and the secretion of BMP-2 protein into the medium. This "coupled" model of bone formation will allow the future investigation of various stimuli on the process of normal bone formation/remodeling as it relates to the cellular function of osteoblasts, osteoclasts and osteocytes in the generation of human mineralized tissue. (C) 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.