Perfused multiwell plate for 3D liver tissue engineering.
Perfused multiwell plate for 3D liver tissue engineering.
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DOI：
10.1039/b913221j
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发表时间：
2010-01-07
期刊：
影响因子：
6.1
通讯作者：
Griffith LG
中科院分区：
文献类型：
--
作者：
Domansky K;Inman W;Serdy J;Dash A;Lim MH;Griffith LG
体外模型以一种可扩展和易于使用的形式捕捉体内组织和器官行为的复杂性，是药物发现所需的。为了解决这个问题，我们开发了一种生物反应器，它可以在持续灌流的情况下维持3D组织培养，我们还将多个生物反应器集成到一个多孔板形式的阵列中。所有的生物反应器都是流体隔离的。阵列中的每个生物反应器包含支持形成数百个3D微尺度组织单元的支架。组织单元通过集成的气动隔膜微泵向生物反应器内循环的细胞培养液灌流。泵的电子控制保持在培养箱外，并通过气动管路连接到灌流的多井。对接设计和开孔生物反应器布局使处理灌流的多孔培养板类似于使用标准的多孔组织培养板。循环培养液中氧的消耗和运输的模型被用来预测原代肝培养的适当操作参数。然后，在培养开始后，测量系统中关键位置的氧气浓度作为流速和时间的函数，以确定氧气消耗率。在培养7天后，通过肝细胞和肝窦内皮细胞(LSEC)表型标记的免疫染色，在灌流的多孔反应器中种植的细胞形成的组织仍然具有功能活性。
In vitro models that capture the complexity of in vivo tissue and organ behaviors in a scalable and easy-to-use format are desirable for drug discovery. To address this, we have developed a bioreactor that fosters maintenance of 3D tissue cultures under constant perfusion and we have integrated multiple bioreactors into an array in a multiwell plate format. All bioreactors are fluidically isolated from each other. Each bioreactor in the array contains a scaffold that supports formation of hundreds of 3D microscale tissue units. The tissue units are perfused with cell culture medium circulated within the bioreactor by integrated pneumatic diaphragm micropumps. Electronic controls for the pumps are kept outside the incubator and connected to the perfused multiwell by pneumatic lines. The docking design and open-well bioreactor layout make handling perfused multiwell plates similar to using standard multiwell tissue culture plates. A model of oxygen consumption and transport in the circulating culture medium was used to predict appropriate operating parameters for primary liver cultures. Oxygen concentrations at key locations in the system were then measured as a function of flow rate and time after initiation of culture to determine oxygen consumption rates. After seven days in culture, tissue formed from cells seeded in the perfused multiwell reactor remained functionally viable as assessed by immunostaining for hepatocyte and liver sinusoidal endothelial cell (LSEC) phenotypic markers.
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影响因子：
--
作者：
MATSUMURA, T;KAUFFMAN, FC;THURMAN, RG
通讯作者：
THURMAN, RG
影响因子：
3.8
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Powers, MJ;Domansky, K;Griffith, LG
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通讯作者：
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