Analysis of Solution Exchange in Flow Chambers with Applications to Cell Membrane Permeability Measurement

作者：Higgins Adam Z; Karlsson Jens O M^*

来源：Cellular and Molecular Bioengineering, 2010, 3(3): 269-285.

DOI：10.1007/s12195-010-0125-z

• Parallel-plate flow chamber
• Fluorescence quenching
• Osmosis
• Water transport
• Hydraulic conductivity
• Beta cell

Cell membrane permeability estimation using flow chamber experiments is susceptible to errors caused by non-negligible solution exchange time after switching of perfusate reservoirs. To prevent such confounding effects, we have undertaken theoretical and experimental analyses of the mass transport of osmotically active solutes. A diffusion-convection model was used to predict the kinetics of solution exchange as a function of Peclet number (Pe) and chamber geometry, yielding guidelines for the design of flow chambers optimized for permeability measurement. Common experimental methods for quantifying solution exchange kinetics (using transmittance or absorbance measurements) were also simulated, and found to be associated with significant error. We therefore used a confocal microscopy technique to validate the dependence of solute exchange kinetics on Pe; the solution exchange time was negligible for flow rates with Pe > 10(6). A fluorescence quenching method was used to estimate the membrane water permeability (L(p)) of mouse insulinoma (MIN6) cells in adherent monolayer cultures, yielding L(p)A/V(w0) = (4.4 +/- 0.1) x 10(-8) Pa(-1) s(-1) (where A/V(w0) is the ratio of cell surface area to isotonic water volume).

• 出版日期2010-9