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Characterization of mouse alveolar epithelial cell monolayers.
Am J Physiol Lung Cell Mol Physiol. 2009 Jun; 296(6):L1051-8.AJ

Abstract

We investigated the influence of extracellular matrix on transport properties of mouse alveolar epithelial cell (AEC) monolayers (MAECM) and transdifferentiation of isolated mouse alveolar epithelial type II (AT2) cells into an alveolar epithelial type I (AT1) cell-like phenotype. Primary mouse AT2 cells plated on laminin 5-coated polycarbonate filters formed monolayers with transepithelial resistance (R(T)) and equivalent short-circuit current (I(EQ)) of 1.8 kOmega.cm(2) and 5.3 microA/cm(2), respectively, after 8 days in culture. Amiloride (10 microM), ouabain (0.1 mM), and pimozide (10 microM) decreased MAECM I(EQ) to 40%, 10%, and 65% of its initial value, respectively. Sequential addition of pimozide and amiloride, in either order, revealed that their inhibitory effects are additive, suggesting that cyclic nucleotide-gated channels contribute to amiloride-insensitive active ion transport across MAECM. Ussing chamber measurements of unidirectional ion fluxes across MAECM under short-circuit conditions indicated that net absorption of Na(+) in the apical-to-basolateral direction is comparable to net ion flux calculated from the observed short-circuit current: 0.38 and 0.33 microeq.cm(-2).h(-1), respectively. Between days 1 and 9 in culture, AEC demonstrated increased expression of aquaporin-5 protein, an AT1 cell marker, and decreased expression of pro-surfactant protein-C protein, an AT2 cell marker, consistent with transition to an AT1 cell-like phenotype. These results demonstrate that AT1 cell-like MAECM grown on laminin 5-coated polycarbonate filters exhibit active and passive transport properties that likely reflect the properties of intact mouse alveolar epithelium. This mouse in vitro model will enhance the study of AEC derived from mutant strains of mice and help define important structure-function correlations.

Authors+Show Affiliations

Department of Medicine, Will Rogers Institute Pulmonary Research Center, University of Southern California, Los Angeles, California, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19329539

Citation

Demaio, Lucas, et al. "Characterization of Mouse Alveolar Epithelial Cell Monolayers." American Journal of Physiology. Lung Cellular and Molecular Physiology, vol. 296, no. 6, 2009, pp. L1051-8.
Demaio L, Tseng W, Balverde Z, et al. Characterization of mouse alveolar epithelial cell monolayers. Am J Physiol Lung Cell Mol Physiol. 2009;296(6):L1051-8.
Demaio, L., Tseng, W., Balverde, Z., Alvarez, J. R., Kim, K. J., Kelley, D. G., Senior, R. M., Crandall, E. D., & Borok, Z. (2009). Characterization of mouse alveolar epithelial cell monolayers. American Journal of Physiology. Lung Cellular and Molecular Physiology, 296(6), L1051-8. https://doi.org/10.1152/ajplung.00021.2009
Demaio L, et al. Characterization of Mouse Alveolar Epithelial Cell Monolayers. Am J Physiol Lung Cell Mol Physiol. 2009;296(6):L1051-8. PubMed PMID: 19329539.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization of mouse alveolar epithelial cell monolayers. AU - Demaio,Lucas, AU - Tseng,Wanru, AU - Balverde,Zerlinde, AU - Alvarez,Juan R, AU - Kim,Kwang-Jin, AU - Kelley,Diane G, AU - Senior,Robert M, AU - Crandall,Edward D, AU - Borok,Zea, Y1 - 2009/03/27/ PY - 2009/3/31/entrez PY - 2009/3/31/pubmed PY - 2009/7/17/medline SP - L1051 EP - 8 JF - American journal of physiology. Lung cellular and molecular physiology JO - Am. J. Physiol. Lung Cell Mol. Physiol. VL - 296 IS - 6 N2 - We investigated the influence of extracellular matrix on transport properties of mouse alveolar epithelial cell (AEC) monolayers (MAECM) and transdifferentiation of isolated mouse alveolar epithelial type II (AT2) cells into an alveolar epithelial type I (AT1) cell-like phenotype. Primary mouse AT2 cells plated on laminin 5-coated polycarbonate filters formed monolayers with transepithelial resistance (R(T)) and equivalent short-circuit current (I(EQ)) of 1.8 kOmega.cm(2) and 5.3 microA/cm(2), respectively, after 8 days in culture. Amiloride (10 microM), ouabain (0.1 mM), and pimozide (10 microM) decreased MAECM I(EQ) to 40%, 10%, and 65% of its initial value, respectively. Sequential addition of pimozide and amiloride, in either order, revealed that their inhibitory effects are additive, suggesting that cyclic nucleotide-gated channels contribute to amiloride-insensitive active ion transport across MAECM. Ussing chamber measurements of unidirectional ion fluxes across MAECM under short-circuit conditions indicated that net absorption of Na(+) in the apical-to-basolateral direction is comparable to net ion flux calculated from the observed short-circuit current: 0.38 and 0.33 microeq.cm(-2).h(-1), respectively. Between days 1 and 9 in culture, AEC demonstrated increased expression of aquaporin-5 protein, an AT1 cell marker, and decreased expression of pro-surfactant protein-C protein, an AT2 cell marker, consistent with transition to an AT1 cell-like phenotype. These results demonstrate that AT1 cell-like MAECM grown on laminin 5-coated polycarbonate filters exhibit active and passive transport properties that likely reflect the properties of intact mouse alveolar epithelium. This mouse in vitro model will enhance the study of AEC derived from mutant strains of mice and help define important structure-function correlations. SN - 1040-0605 UR - https://www.unboundmedicine.com/medline/citation/19329539/Characterization_of_mouse_alveolar_epithelial_cell_monolayers_ L2 - http://journals.physiology.org/doi/full/10.1152/ajplung.00021.2009?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -