Abstract
AbstractThe measurement of transepithelial electrical resistance (TEER) is a common technique to determine the barrier integrity of epithelial cell monolayers. However, it is remarkable that absolute TEER values of similar cell types cultured under comparable conditions show an immense heterogeneity. Based on previous observations, we hypothesized that the heterogeneity of absolute TEER measurements can not only be explained by maturation of junctional proteins but rather by dynamics in the absolute length of cell junctions within monolayers. Therefore, we analyzed TEER in epithelial cell monolayers of Caco2 cells during their differentiation, with special emphasis on both changes in the junctional complex and overall cell morphology within monolayers. We found that in epithelial Caco2 monolayers TEER increased until confluency, then decreased for some time, which was then followed by an additional increase during junctional differentiation. In contrast, permeability of macromolecules measured at different time points as 4 kDA fluorescein isothiocyanate (FITC)-dextran flux across monolayers steadily decreased during this time. Detailed analysis suggested that this observation could be explained by alterations of junctional length along the cell borders within monolayers during differentiation. In conclusion, these observations confirmed that changes in cell numbers and consecutive increase of junctional length have a critical impact on TEER values, especially at stages of early confluency when junctions are immature.
Funder
Deutsche Forschungsgesellschaft
DFG priority program
Interdisziplinäres Zentrum für Klinische Forschung, Universitätsklinikum Würzburg
Universitätsklinikum Würzburg
Publisher
Springer Science and Business Media LLC
Subject
Cell Biology,Medical Laboratory Technology,Molecular Biology,Histology
Reference23 articles.
1. Blume LF, Denker M, Gieseler F, Kunze T (2010) Temperature corrected transepithelial electrical resistance (TEER) measurement to quantify rapid changes in paracellular permeability. Pharmazie 65(1):19–24
2. Bowman PD, Ennis SR, Rarey KE, Betz AL, Goldstein GW (1983) Brain microvessel endothelial cells in tissue culture: a model for study of blood-brain barrier permeability. Ann Neurol 14(4):396–402. https://doi.org/10.1002/ana.410140403
3. Briske-Anderson MJ, Finley JW, Newman SM (1997) The influence of culture time and passage number on the morphological and physiological development of Caco-2 cells. Proc Soc Exp Biol Med 214(3):248–257. https://doi.org/10.3181/00379727-214-44093
4. Chang YS, Munn LL, Hillsley MV, Dull RO, Yuan J, Lakshminarayanan S, Gardner TW, Jain RK, Tarbell JM (2000) Effect of vascular endothelial growth factor on cultured endothelial cell monolayer transport properties. Microvasc Res 59(2):265–277. https://doi.org/10.1006/mvre.1999.2225
5. Duffy SL, Murphy JT (2001) Colorimetric assay to quantify macromolecule diffusion across endothelial monolayers. Biotechniques 31(3):495–496. https://doi.org/10.2144/01313st02 (PMID: 11570492)
Cited by
28 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献