A chip-based platform for the in vitro generation of tissues in three-dimensional organization
A chip-based platform for the in vitro generation of tissues in three-dimensional organization
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DOI：
10.1039/b618488j
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发表时间：
2007-06-01
期刊：
影响因子：
6.1
通讯作者：
Weibezahn, Karl-Friedrich
中科院分区：
文献类型：
--
作者：
Gottwald, Eric;Giselbrecht, Stefan;Weibezahn, Karl-Friedrich
我们描述了一个用于组织三维培养的多功能平台。该装置由具有 1-2 cm(2) 微结构面积的聚合物芯片组成。该芯片要么被构造为尺寸为 120-300 x 300 x 300 Pm（高 x 1 x 宽）的微型容器网格，要么被构造为圆形凹槽阵列（直径 300 μm，深 300 μm）。微容器可以单独配备可寻址的 3D 微电极，从而可以对可兴奋细胞进行电刺激并现场测量电化学可访问的参数。该系统适用于高达 8 x 1 06 个细胞的高细胞密度培养，并且由于矩形网格布局，允许对培养细胞进行自动显微镜分析。超过 1000 个微容器能够在超灌注/灌注条件下并行分析不同参数。通过将不同的聚合物芯片与各种类型的生物反应器结合使用，我们证明了基于芯片的生物反应器对于组织培养应用的主要适用性。原代细胞系和已建立的细胞系已成功培养并分析其功能特性。当细胞在非灌注芯片中培养时，随着时间的推移，可以观察到相当程度的细胞凋亡，表明需要主动灌注。这里介绍的系统也已应用于多能胚胎干细胞的分化分析，并且可能适用于干细胞生态位的分析。
We describe a multi-purpose platform for the three-dimensional cultivation of tissues. The device is composed of polymer chips featuring a microstructured area of 1-2 cm(2). The chip is constructed either as a grid of micro-containers measuring 120-300 x 300 x 300 Pm (h x 1 x w), or as an array of round recesses (300 mu m diameter, 300 mu m deep). The micro-containers may be separately equipped with addressable 3D-micro-electrodes, which allow for electrical stimulation of excitable cells and on-site measurements of electrochemically accessible parameters. The system is applicable for the cultivation of high cell densities of up to 8 x 1 06 cells and, because of the rectangular grid layout, allows the automated microscopical analysis of cultivated cells. More than 1000 micro-containers enable the parallel analysis of different parameters under superfusion/perfusion conditions. Using different polymer chips in combination with various types of bioreactors we demonstrated the principal suitability of the chip-based bioreactor for tissue culture applications. Primary and established cell lines have been successfully cultivated and analysed for functional properties. When cells were cultured in non-perfused chips, over time a considerable degree of apoptosis could be observed indicating the need for an active perfusion. The system presented here has also been applied for the differentiation analysis of pluripotent embryonic stem cells and may be suitable for the analysis of the stem cell niche.