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Microchamber device for detection of transporter activity of adherent cells.

Tsugane M, Uejima E, Suzuki H - Front Bioeng Biotechnol (2015)

Bottom Line: When adherent cells are seeded onto the poly-di-methyl siloxane 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 over-expressing MDR1, an ATP-binding cassette transporter, and succeeded in detecting the transport of fluorescence-conjugated paclitaxel, the anti-cancer drug, at the single-cell level.

View Article: PubMed Central - PubMed

Affiliation: Department of Precision Mechanics, Faculty of Science and Engineering, Chuo University , Tokyo , Japan ; Department of Clinical Pharmacy Research and Education, Graduate School of Pharmaceutical Sciences, Osaka University , Osaka , Japan.

ABSTRACT
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 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 over-expressing MDR1, an ATP-binding cassette transporter, and succeeded in detecting the transport of fluorescence-conjugated paclitaxel, the anti-cancer drug, at the single-cell level.

No MeSH data available.


Schematic of the microchamber system for detection of transporter activity expressed in adherent cells. The efflux of anti-cancer drug accumulates in the microchamber sealed by the intact cells for detection by imaging.
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Figure 1: Schematic of the microchamber system for detection of transporter activity expressed in adherent cells. The efflux of anti-cancer drug accumulates in the microchamber sealed by the intact cells for detection by imaging.

Mentions: In the pharmacotherapy of cancer, there is wide variation among individuals in treatment efficacy and the development of side effects from anti-cancer drugs, due to the acquisition of resistance by cancer cells as well as polymorphisms in patients. A clinical diagnostic device that detects the activity of transporters based on cells collected from cancer patient biopsies could help predict the efficacy and side effects of certain drugs, and contribute to the selection of appropriate anti-cancer treatments for individual patients. As seen in the above examples, transport of substances via membrane transporters can be detected rapidly and easily at the single-cell or single-molecule level using microdevices. As solid tumors account for most tumors that consist of adherent cells, a microdevice that measures transport activity of adherent cells under near-physiological conditions is of particular importance. Thus, in this study, we develop a microchamber device for the detection of transport activity, in which drugs, as substrates, are exported from cells and accumulate within a confined space of the microchamber directly covered by adherent, cultured cells (Figure 1). Here, cultured cells over-expressing MDR1 proteins were used to test the performance of the constructed microchamber device in measuring the transport activity of anti-cancer drugs.


Microchamber device for detection of transporter activity of adherent cells.

Tsugane M, Uejima E, Suzuki H - Front Bioeng Biotechnol (2015)

Schematic of the microchamber system for detection of transporter activity expressed in adherent cells. The efflux of anti-cancer drug accumulates in the microchamber sealed by the intact cells for detection by imaging.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4364289&req=5

Figure 1: Schematic of the microchamber system for detection of transporter activity expressed in adherent cells. The efflux of anti-cancer drug accumulates in the microchamber sealed by the intact cells for detection by imaging.
Mentions: In the pharmacotherapy of cancer, there is wide variation among individuals in treatment efficacy and the development of side effects from anti-cancer drugs, due to the acquisition of resistance by cancer cells as well as polymorphisms in patients. A clinical diagnostic device that detects the activity of transporters based on cells collected from cancer patient biopsies could help predict the efficacy and side effects of certain drugs, and contribute to the selection of appropriate anti-cancer treatments for individual patients. As seen in the above examples, transport of substances via membrane transporters can be detected rapidly and easily at the single-cell or single-molecule level using microdevices. As solid tumors account for most tumors that consist of adherent cells, a microdevice that measures transport activity of adherent cells under near-physiological conditions is of particular importance. Thus, in this study, we develop a microchamber device for the detection of transport activity, in which drugs, as substrates, are exported from cells and accumulate within a confined space of the microchamber directly covered by adherent, cultured cells (Figure 1). Here, cultured cells over-expressing MDR1 proteins were used to test the performance of the constructed microchamber device in measuring the transport activity of anti-cancer drugs.

Bottom Line: When adherent cells are seeded onto the poly-di-methyl siloxane 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 over-expressing MDR1, an ATP-binding cassette transporter, and succeeded in detecting the transport of fluorescence-conjugated paclitaxel, the anti-cancer drug, at the single-cell level.

View Article: PubMed Central - PubMed

Affiliation: Department of Precision Mechanics, Faculty of Science and Engineering, Chuo University , Tokyo , Japan ; Department of Clinical Pharmacy Research and Education, Graduate School of Pharmaceutical Sciences, Osaka University , Osaka , Japan.

ABSTRACT
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 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 over-expressing MDR1, an ATP-binding cassette transporter, and succeeded in detecting the transport of fluorescence-conjugated paclitaxel, the anti-cancer drug, at the single-cell level.

No MeSH data available.