一次性多室微细胞培养装置的研制

H. Nakayama, H. Kimura, M. Nishikawa, K. Komori, T. Fujii, Y. Sakai
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引用次数: 1

摘要

我们的最终目标是开发一种“片上动物/人”的多室微细胞培养系统。为了实现最终目标,我们设计并开发了一种一次性的三室微细胞培养,使用基于聚二甲基硅氧烷(PDMS)的微加工和基于小型磁力搅拌器的内部泵送系统(用于一次性和节省空间)。所开发的装置能够在两种不同的灌注模式下运行,即每室灌注模式和整个装置灌注模式,从而可以在不同的培养方案下生长或维持来自不同组织/器官的细胞,最后将它们连接起来进行整个装置的毒性动力学研究。为了评估化学分布和生物代谢过程,我们在设备中加入了脂肪和肝组织来源的细胞,因为它们控制疏水和致癌化学物质的分布和生物转化。肝源性细胞,人肝癌hepg2,被接种并培养在肝室和脂肪组织室中,其中成熟的大鼠脂肪细胞被固定在3D支架中。在灌注过程中,两种细胞均以单层形式附着在每个隔室的底表面,没有脱离或形成漂浮聚集体。大鼠原代肝细胞也在肝组织室中稳定培养。我们现在正在尝试用荧光技术可视化肝室的毒性动力学过程,在使用荧光和致癌化学物质的设备中,在存在和不存在大鼠组织室的情况下。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Development of a disposable multi-compartment micro-cell culture device
Our final goal is development of a multi-compartment micro-cell culture system "on-chip animal/human". Towards the final goal, we designed and developed a disposable-type three-compartment micro-cell culture using polydimethylsiloxane (PDMS)-based microfabrication and small magnetic stirrer-based internal pumping system (for disposability and space-saving). The developed device was able to be operated in two different perfusion modes, that is, an each compartment perfusion mode and an entire device perfusion mode, so that cells derived from different tissue/organ can be grown or maintained in different culture protocols and finally they are connected for toxicokinetic studies over the entire device. To evaluate the chemical distribution and biological metabolic processes, we incorporated fat and liver-tissue derived cells in the device because they control the distribution and biotransformation of hydrophobic and carcinogenic chemicals. Liver-derived cells, human hepatocarcinoma Hep G2, were inoculated and cultured in the liver compartment in addition to the fat tissue compartment where mature rat adipocytes were immobilized in a 3D scaffold. Both cells remained attached to the surface in monolayers on the bottom surfaces of each compartment without detachment or forming floating aggregates during perfusion. Rat primary hepatocytes were also stably cultured in the liver tissue compartment. We are now trying fluorescent-based visualization of toxicokinetic processes in the liver compartment in the present and absence of rat tissue compartment in the devices administered with fluorescent and carcinogenic chemicals.
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