Microbioassay system for antiallergic drug screening using suspension cells retaining in a poly(dimethylsiloxane) microfluidic device
Microbioassay system for antiallergic drug screening using suspension cells retaining in a poly(dimethylsiloxane) microfluidic device
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DOI：
10.1021/ac048288o
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发表时间：
2005-05-15
影响因子：
7.4
通讯作者：
Okubo, A
中科院分区：
文献类型：
--
作者：
Tokuyama, T;Fujii, S;Okubo, A
本文介绍了一种在多层微芯片上检测肥大细胞（悬浮细胞）释放的组胺的抗过敏药物筛选系统。本研究采用弹性材料聚二甲基硅氧烷（PDMS）制备微通道和微室。微芯片由两个部分组成：一个是组胺释放部分，它有一个细胞室，另一个是组胺衍生部分。两者都被层压在一个微芯片上。用硝酸纤维素膜过滤系统将大鼠腹膜肥大细胞保留在细胞室（1.2 μ L）中。这种过滤系统可以很容易地保留悬浮细胞而不损坏细胞。肥大细胞在足够的时间内存活，以便在细胞室上进行试验。用化学释放化合物48/80 （C48/80）刺激细胞，然后组胺流入下层，在那里与邻苯二醛衍生成荧光分子，并在微芯片上检测其荧光。该流动系统可以检测组胺释放的时间过程，该微芯片系统仅需20分钟。通过这个集成系统，从500个细胞中检测出51 pmol的组胺，与传统的批量系统相比，检测所需的细胞数量减少到1%。通过比较用药前后的组胺释放水平，评价药物的作用。抗过敏药物克莫甘酸二钠（DSCG）对C48/80诱导的组胺释放的抑制率与DSCG浓度有关。该流动系统适用于抗过敏药物筛选，通过快速测量从非常少量的肥大细胞中抑制组胺释放。
This article describes an antiallergic drug-screening system by the detection of histamine released from mast cells (suspension cells) on a multilayer microchip. In this study, the elastmeric material, poly(dimethylsiloxane) (PDMS), was employed to fabricate microchannels and microchambers. The microchip consists of two sections: a histamine-releasing one, which has a cell chamber, and a histamine-derivatizing one. Both were laminated to one microchip. Rat peritoneal mast cells were retained in the cell chamber (1.2 mu L) with a filtering system using a cellulose nitrate membrane. This filtering system could easily retain suspension cells without cell damage. Mast cells were viable for a sufficient time to conduct the assay on the cell chamber. The cells were stimulated with a chemical release compound 48/80 (C48/80), and then histamine flowed into the lower layer, where it was derivatized to the fluorescent molecules with o-phthalaldehyde and its fluorescence was detected on the microchip. This flow system could detect the time course of the histamine release, and this microchip system required only 20 min for the assay. By this integrated system, 51 pmol of histamine released from 500 cells was detected, and the number of cells required for the assay was reduced to 1% compared with conventional bulk systems. By comparing the released histamine levels with and without drugs, their effect could be evaluated. The inhibition ratio of C48/80 induced-histamine release using an antiallergic drug, disodium cromoglicate (DSCG), was related to the concentration of DSCG. This flow system was applicable for antiallergy drug screening by rapid measurement of the inhibition of histamine release from a very small amount of mast cells.