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. 2020 Oct 11;12(10):1151. doi: 10.3390/v12101151

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Characterization of NAEPCs in organotypic 3D air–liquid interface (ALI) cell cultures by electron microscopic imaging. (a) NAEPCs were used for 3D ALI cell cultures, were treated with HDM using different concentrations indicating time points, and were imaged with raster-electron microscopy. An irritated epithelium after HDM stimulation with 100 µg/mL was observed. Some cells were also detached from the cell population in the specimens of the asthmatic group. After day 3, we observed an irritation of the epithelium and an affected barrier integrity in some regions of the cultures (scale bars were added to the respective figures, different magnifications were used (500-fold to 2500-fold)). (b) Measuring the thickness of the cilia, there was a lack of significant correlations between treated and untreated, healthy, or asthmatic samples (scale bars were added to the respective figures, different magnifications were used (500× to 2500×)). (c) Histologically, the epithelium was affected and numerous mucus cells were observed when treated with high dose HDM (10× magnification). (d,e) Transmission electron microscopic analyses of untreated healthy (d) and asthmatic (e) 3D cultures revealed a different tight junction conformation, especially for asthmatic 3D cultures. The tight junctions in untreated asthmatic samples (e) were tightly packed and were present in a higher numbers compared to untreated control samples (d). The white arrows mark the respective tight junctions. Different magnifications were applied (20,000-fold to 50,000-fold).

Characterization of NAEPCs in organotypic 3D air–liquid interface (ALI) cell cultures by electron microscopic imaging. (a) NAEPCs were used for 3D ALI cell cultures, were treated with HDM using different concentrations indicating time points, and were imaged with raster-electron microscopy. An irritated epithelium after HDM stimulation with 100 µg/mL was observed. Some cells were also detached from the cell population in the specimens of the asthmatic group. After day 3, we observed an irritation of the epithelium and an affected barrier integrity in some regions of the cultures (scale bars were added to the respective figures, different magnifications were used (500-fold to 2500-fold)). (b) Measuring the thickness of the cilia, there was a lack of significant correlations between treated and untreated, healthy, or asthmatic samples (scale bars were added to the respective figures, different magnifications were used (500× to 2500×)). (c) Histologically, the epithelium was affected and numerous mucus cells were observed when treated with high dose HDM (10× magnification). (d,e) Transmission electron microscopic analyses of untreated healthy (d) and asthmatic (e) 3D cultures revealed a different tight junction conformation, especially for asthmatic 3D cultures. The tight junctions in untreated asthmatic samples (e) were tightly packed and were present in a higher numbers compared to untreated control samples (d). The white arrows mark the respective tight junctions. Different magnifications were applied (20,000-fold to 50,000-fold).