Unstirred Water Layers and the Kinetics of Organic Cation Transport.

ABSTRACT:

Purpose

Unstirred water layers (UWLs) present an unavoidable complication to the measurement of transport kinetics in cultured cells, and the high rates of transport achieved by overexpressing heterologous transporters exacerbate the UWL effect. This study examined the correlation between measured Jmax and Kt values and the effect of manipulating UWL thickness or transport Jmax on the accuracy of experimentally determined kinetics of the multidrug transporters, OCT2 and MATE1.

Methods

Transport of TEA and MPP was measured in CHO cells that stably expressed human OCT2 or MATE1. UWL thickness was manipulated by vigorous reciprocal shaking. Several methods were used to manipulate maximal transport rates.

Results

Vigorous stirring stimulated uptake of OCT2-mediated transp

SUBMITTER: Shibayama T 

PROVIDER: S-EPMC4529401 | biostudies-literature | 2015 Sep

REPOSITORIES: biostudies-literature