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首页> 外文期刊>Acta biomaterialia >A three-dimensional tissue culture model of bone formation utilizing rotational co-culture of human adult osteoblasts and osteoclasts.
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A three-dimensional tissue culture model of bone formation utilizing rotational co-culture of human adult osteoblasts and osteoclasts.

机译：利用成人成骨细胞和破骨细胞的旋转共培养，形成骨骼形成的三维组织培养模型。

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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 (>4mm 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.
机译：活骨是一种复杂的三维复合材料，包括在矿化的细胞外基质中空间排列的多种细胞类型。迄今为止，由于缺乏合适的捕获“耦合”本质的体外模型，阻碍了对参与骨骼对机械和生化刺激反应的主要成骨细胞之间复杂细胞水平串扰的机理研究。这个回应。使用新颖的旋转共培养方法，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生成了大尺寸（直径大于4mm）的三维矿化组织，而无需任何可能干扰其的外源性骨传导支架材料这样的细胞间相互作用。成熟，分化的骨结构由破骨细胞和成骨细胞所占据的外部区域以及包含被包裹在自组装的多孔矿化细胞外基质中的骨细胞的中央区域组成。骨构建物表现出与重塑小梁骨相似的形态，矿物质和生化特征，包括构建物中SOST，BGLAP，ACP5，BMP-2，BMP-4和BMP-7的mRNA表达以及BMP-2蛋白的分泌进入媒体。这种“耦合”的骨形成模型将允许将来研究正常骨形成/重塑过程中的各种刺激，因为它与成骨细胞，破骨细胞和骨细胞在人类矿化组织的生成中的细胞功能有关。

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