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Cell division does not increase transepithelial permeability of LLC-PK1 cell sheets
D T Saladik1, A P Soler, S A Lewis
1Lankenau Medical Research Center, Wynnewood, Pennsylvania 19096, USA.
Experimental Cell Research
|October 1, 1995
Summary
Transepithelial electrical resistance (TER) in LLC-PK1 cells stabilizes after 4 days. Epidermal Growth Factor (EGF) treatment increases cell density and TER by altering paracellular ion permeability, not tight junction size selectivity.
Area of Science:
- Cell Biology
- Biophysics
- Physiology
Background:
- Transepithelial electrical resistance (TER) in LLC-PK1 cell sheets is initially unstable postseeding.
- TER fluctuations require detailed characterization for reliable cell culture models.
Purpose of the Study:
- To investigate the characteristics of TER fluctuations in LLC-PK1 cells after plating.
- To determine the effect of epidermal growth factor (EGF) on TER and associated cellular changes.
Main Methods:
- Monitoring TER and [3H]-thymidine incorporation in LLC-PK1 cell sheets over several days.
- Applying EGF treatment at 24 hours postplating and assessing its impact on TER, cell density, and permeability.
- Measuring transepithelial fluxes of [14C]D-mannitol and conducting diffusion/bi-ionic potential studies.
Main Results:
- TER stabilized around day 4, with peak cell cycle activity preceding peak TER.
- EGF treatment at 24 hours significantly increased TER and cell density by day 4.
- EGF altered charge selectivity of the paracellular pathway, decreasing permeability to Na+ and Cl-, particularly Cl-.
Conclusions:
- TER in LLC-PK1 cell sheets requires 4 days to stabilize postseeding.
- Increased cell density correlates with elevated TER due to altered paracellular ion permeability.
- EGF modulates LLC-PK1 cell sheet ion selectivity, impacting halide anion permeability.