Microbioassay System for Antiallergic Drug Screening Using Suspension Cells Retaining in a Poly(dimethylsiloxane) Microfluidic Device

摘要

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 mat6rial, 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 onemicrochip. Rat peritoneal mast cells were retained in the cell chamber (1.2 PL) with a filtering system using acellulose nitrate membrane. This filtering system could easily retain suspension cells without cell damage. Mastcells were viable for a sufficient time to conduct the assay on the cell chamber. The cells were stimulated with achemical release compound 48/80 (C48/80), and then histamine flowed into the lower farer, where it was derivatized to the nuorescent molecules with o-phthalal-dehyde and its fluorescence was detected on the microchip. This now system could detect the time course of the histamine release, and this microchip system required only 20 min for the assay. By this nitegrated system, 51 pmol of histamine released from 500 cells was detected, and the number of cells required for the assay was reduced to 1percent 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.