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Updated: Aug 17, 2026
Models and Methods to Evaluate Transport of Drug Delivery Systems Across Cellular Barriers
Published on: October 17, 2013
The Madin Darby canine kidney (MDCK) epithelial cell monolayer as a model cellular transport barrier
M J Cho1, D P Thompson, C T Cramer
1Pharmaceutical Research and Development Division, Upjohn Company, Kalamazoo, Michigan 49001.
Abstract:
Two strains of Madin Darby canine kidney (MDCK) cells were grown on a polycarbonate membrane with 3-micron pores without any extracellular matrix treatment. The membrane, 2.45 cm in diameter, which is part of a commercially obtained presterilized culture insert, provides two chambers when placed in a regular six-well culture plate. This device was found to be convenient for investigating transport of a few selected fluid-phase markers across the MDCK cell monolayer. Both the strain from the American Type Culture Collection (ATCC) and the so-called highly resistant strain I, at a serial passage between 65 and 70, showed a seeding concentration-dependent lag phase followed by a growth phase with a 21-hr doubling time. When seeded at 5 x 10(4) cells/cm2, cell confluence was achieved in 5 days in a modified Eagle's minimum essential medium (MEM) containing 10% fetal bovine serum under a 5% CO2 atmosphere. Similarly, transepithelial electrical resistance (TEER) also reached a plateau value in 5 days. Both light and electron microscopic examinations revealed well-defined junctional structures. Transport of the fluid-phase markers, sucrose, lucifer yellow CH (LY), inulin, and dextran across the MDCK cell monolayers was studied primarily at 37 degrees C following the apical-to-basolateral as well as the basolateral-to-apical direction. Large variations in the steady-state transport rate were observed for a given marker between the cell layer preparations. Thus, the present study proposes an "internal standard" procedure for meaningful comparisons of the transport rate. When normalized to the rate of sucrose, the rate ratio was 1.00:0.80:0.67:0.15 for sucrose:LY:inulin:dextran.(ABSTRACT TRUNCATED AT 250 WORDS)
Insights
This study developed a novel "internal standard" method to accurately compare fluid-phase marker transport rates across Madin Darby canine kidney (MDCK) cell monolayers, improving research reliability.
Area of Science:
- Cell Biology
- Biophysics
- Pharmacology
Background:
- Madin Darby canine kidney (MDCK) cells are a standard model for studying epithelial transport.
- Investigating transport across cell monolayers requires consistent and reproducible methods.
- Variability in transport rates can hinder accurate comparisons between experiments.
Purpose of the Study:
- To establish a reliable method for measuring fluid-phase marker transport across MDCK cell monolayers.
- To introduce an "internal standard" procedure for normalizing transport rates.
- To compare the transport rates of various fluid-phase markers.
Main Methods:
- Culturing two strains of MDCK cells on polycarbonate membranes without extracellular matrix.
- Utilizing a two-chamber culture insert system for transport studies.
- Measuring transepithelial electrical resistance (TEER) and observing cell morphology via light and electron microscopy.
- Quantifying the transport of sucrose, lucifer yellow CH (LY), inulin, and dextran in both directions across the monolayer.
Main Results:
- MDCK cell monolayers reached confluence and stable transepithelial electrical resistance (TEER) within 5 days.
- Well-defined junctional structures were observed in both light and electron microscopy.
- Significant variations in steady-state transport rates were noted between cell preparations.
- A normalized transport rate ratio (sucrose:LY:inulin:dextran) of 1.00:0.80:0.67:0.15 was established using sucrose as the internal standard.
Conclusions:
- The developed method provides a reproducible system for studying MDCK cell monolayer transport.
- The proposed "internal standard" procedure allows for meaningful comparisons of marker transport rates.
- This approach enhances the reliability and accuracy of epithelial transport research.
