Microchamber Device for Detection of Transporter Activity of Adherent Cells

• Main Authors: Mamiko eTsugane, Etsuko eUejima, Hiroaki eSUZUKI
• Format: Article
• Language: English
• Published: Frontiers Media S.A. 2015-03-01
• Series: Frontiers in Bioengineering and Biotechnology
• Subjects: transporter; Anti-cancer drug; Microchamber; Adherent cell; Transport Assay
• Online Access: http://journal.frontiersin.org/Journal/10.3389/fbioe.2015.00032/full

We present a method to detect the transporter activity of intact adherent cells using a microchamber device. When adherent cells are seeded onto the poly-di-methyl siloxane (PDMS) substrate having microchambers with openings smaller than the size of a cell, the cells form a confluent layer that covers the microchambers, creating minute, confined spaces. As substances exported across the cell membrane accumulate, transporter activity can be detected by observing the fluorescence intensity increase in the microchamber. We tested the microchamber device with HeLa cells overexpressing MDR1, an ATP-binding cassette (ABC) transporter, and succeeded in detecting the transport of fluorescence-conjugated paclitaxel, the anti-cancer drug, at the single-cell level.