A continuous perfusion microplate for cell culture
A continuous perfusion microplate for cell culture
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DOI：
10.1039/c2lc41102d
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发表时间：
2013-01-01
期刊：
影响因子：
6.1
通讯作者：
Yuen, Po Ki
中科院分区：
文献类型：
--
作者：
Goral, Vasiliy N.;Zhou, Chunfeng;Yuen, Po Ki
我们描述了一种具有流体连接的威尔斯孔的96孔微孔板，其能够在威尔斯孔之间进行连续的流体灌注，而不需要外部泵送。这种灌注微孔板中的单个单元由三个威尔斯孔组成：源孔、中间的样品（细胞培养物）孔和废物孔。流体灌注是利用由威尔斯孔中的不同液位产生的流体静压和通过连接威尔斯孔的纤维素膜的窄条芯吸的流体的组合来实现的。有一个很好的对应关系之间所观察到的灌注流动力学和基于达西定律的流动模拟。在灌注微孔板中培养4天且细胞培养孔中未更换培养基的肝细胞（C3 A细胞）与暴露于每日更换培养基的肝细胞具有相同的活力。需要由LADMAC细胞调节的培养基的EOC 20细胞显示在灌注微孔板的相邻细胞培养孔中与在源孔中培养的LADMAC细胞具有相同的活力。当将替加氟（一种前药）加入源孔中的原代人肝细胞中时，其代谢为细胞毒性代谢产物，可杀死在相邻细胞培养孔中培养的结肠癌细胞（HCT 116）;当源孔中仅存在培养基时，未观察到毒性。这些结果表明，灌注微孔板是一个有用的工具，用于各种细胞培养应用的好处，从节省劳动力，使体内毒性研究。
We describe a 96-well microplate with fluidically connected wells that enables the continuous fluid perfusion between wells without the need for external pumping. A single unit in such a perfusion microplate consists of three wells: a source well, a sample ( cell culture) well in the middle and a waste well. Fluid perfusion is achieved using a combination of the hydrostatic pressure generated by different liquid levels in the wells and the fluid wicking through narrow strips of a cellulose membrane connecting the wells. There is an excellent correspondence between the observed perfusion flow dynamics and the flow simulations based on Darcy's Law. Hepatocytes (C3A cells) cultured for 4 days in the perfusion microplate with no media exchange in the cell culture well had the same viability as hepatocytes exposed to a daily exchange of media. EOC 20 cells that require media conditioned by LADMAC cells were shown to be equally viable in the adjacent cell culture well of the perfusion microplate with LADMAC cells cultured in the source well. Tegafur, a prodrug, when added to primary human hepatocytes in the source well, was metabolized into a cytotoxic metabolite that kills colon cancer cells (HCT 116) cultured in the adjacent cell culture well; no toxicity was observed when only medium was in the source well. These results suggest that the perfusion microplate is a useful tool for a variety of cell culture applications with benefits ranging from labor savings to enabling in vivo-like toxicity studies.