Secretory cell types and cell proliferation of human bronchial epithelial cells in an organ-culture system

Bals, Robert; Gamarra, Fernando; Kaps, Anna; Grundler, Stefan; Huber, Rudolf M.; Welsch, Ulrich

doi:10.1007/s004410051150

Cited by 17 publications

(8 citation statements)

References 27 publications

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“…These well-differentiated spheroidlike structures of human airway epithelial cells could be maintained for up to 2 months in suspension culture. We could identify here, after 15 days of 3-D culture, secretory cells typical of goblet cells with secretory granules as described by Bals et al (1998) in organ-culture system of bronchial epithelial cells. Lesuffleur et al (1993) previously reported that mucin secretion is related to the growth and differentiation of different mucus-secreting intestinal epithelial cells.…”

Section: Discussionsupporting

confidence: 67%

Polarized Expression of Cystic Fibrosis Transmembrane Conductance Regulator and Associated Epithelial Proteins during the Regeneration of Human Airway Surface Epithelium in Three-Dimensional Culture

Castillon

Hinnrasky

Zahm

et al. 2002

Laboratory Investigation

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SUMMARY:We have previously shown that, in normal human airway tissue, localization of the cystic fibrosis transmembrane conductance regulator (CFTR) can be affected by epithelial maturation, polarity, and differentiation and that CFTR trafficking and apical localization depend on the integrity of the airway epithelium. In this study, we addressed the question of whether the three-dimensional (3-D) organization of adult human airway epithelial cells in suspension culture under rotation, leading to spheroid-like structures, could mimic the in vivo phenomenon of differentiation and polarization. The kinetics of the differentiation, polarity, and formation of the CFTR-ZO-1-ezrin complex was analyzed by transmission, scanning, and immunofluorescence microscopy. Functional activity of the airway surface epithelium was assessed by monitoring the degree of cAMP-stimulated chloride efflux from cultured cells. Our results show that after the initial step of dedifferentiation, characterized by a loss of ciliated cells and disappearance of epithelial subapical CFTR-ezrin-ZO-1 complex, the isolated cells formed 3-D spheroid structures within 24 hours. After 15 days, progressive ciliogenesis was observed and secretory cells could be identified. After 35 days of 3-D culture, ZO-1, CFTR, ezrin, and CD59 were apically or subapically located, and well-differentiated secretory and ciliated cells were identified. CFTR functionality was assessed by analyzing the Cl Ϫ secretion after amiloride and forskolin perfusion. After 35 days of culture of spheroids in suspension, a significant increase in Cl Ϫ efflux was observed in well-differentiated ciliated cells. (Lab Invest 2002, 82:989 -998).

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Section: Discussionsupporting

confidence: 67%

Polarized Expression of Cystic Fibrosis Transmembrane Conductance Regulator and Associated Epithelial Proteins during the Regeneration of Human Airway Surface Epithelium in Three-Dimensional Culture

Castillon

Hinnrasky

Zahm

et al. 2002

Laboratory Investigation

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“…DOI 10.1002/em cells in the majority of studies [Lin and Chang, 2008]. Since the three-dimensional (3D) architecture of the tissue of origin is lost in these culture conditions, some authors prefer using different organ culture methods to simulate the in vivo situation in a superior way [Bals et al, 1998]. Inspired by techniques creating organ cultures from adenoid tissue [Steinsvåg et al, 1991], 3D nasal mucosa MOCs, which were derived from human inferior turbinate tissue samples, were established [Kleinsasser et al, 2001].…”

Section: Discussionmentioning

confidence: 99%

Repetitive exposure to zinc oxide nanoparticles induces dna damage in human nasal mucosa mini organ cultures

Hackenberg

Zimmermann

Scherzed

et al. 2011

Environ and Mol Mutagen

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Data on the toxicological properties of zinc oxide nanoparticles (ZnO-NPs) is incomplete. ZnO-NPs may enter humans via inhalation or ingestion. The aim of the current study was to evaluate ZnO-NP-induced genotoxicity in three-dimensional (3D) mini organ cultures (MOCs) of human nasal mucosa following repeated exposure to ZnO-NP and regeneration. Nasal MOCs of 10 patients and ZnO-NPs were cultivated for one week and then characterized by electron microscopy. Nasal MOCs were partially covered by ciliated epithelium after one week of cultivation. ZnO-NPs were distributed to the cytoplasm and the nucleus. MOCs were exposed once, twice, or three times to 0.1 or 5 μg/ml of ZnO-NPs for 1 hr per exposure and were then evaluated for cytotoxicity and genotoxicity. MOCs were cultivated for 24 hr after the triple ZnO-NP exposure to allow for regeneration. ZnO-NP exposure did not result in significant cytotoxicity or apoptosis, as determined by trypan blue exclusion and caspase-3 activity, respectively. A significant increase in DNA damage was detected following repetitive exposure compared to single exposure to ZnO-NPs at 5 μg/ml, but not 0.1 μg/ml ZnO-NPs. At both concentrations of ZnO-NP, DNA fragmentation increased after 24 hr of regeneration. In contrast, DNA damage which was induced by the positive control, methyl methanesulfonate, was significantly reduced after 24-hr regeneration. Thus, our results suggest that repetitive exposure to low concentrations of ZnO-NPs results in persistent or ongoing DNA damage.

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“…Thus the three-dimensional organ culture system consists of a nucleus of connective tissue which is completely covered by a multilayered respiratory epithelium. The cultured tissue contains a basal membrane, ciliated cells, secretory cells and basal cells [12]and strongly resembles respiratory epithelium in vivo.…”

Section: Methodsmentioning

confidence: 99%

Kinetics of 5-Aminolevulinic Acid-Induced Fluorescence in Organ Cultures of Bronchial Epithelium and Tumor

et al. 2002

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Background: 5-Aminolevulinic acid (5-ALA)-induced protoporphyrin IX (PPIX) fluorescence improves the differentiation of tumor and normal tissue in the bladder, skin and brain. Objective: The kinetics of 5-ALA-induced protoporphyrin IX (PPIX) fluorescence in organ cultures of normal human bronchial epithelium and cocultures of bronchial epithelium and tumor have been studied. Methods: Cultured biopsies of bronchial epithelium were exposed for 5 or 15 min, or continuously to 5-ALA. PPIX fluorescence was quantified for up to 300 min by spectroscopy. Cocultures of normal bronchial epithelium and a non-small-cell lung cancer cell line (EPLC-32M1) were incubated with 5-ALA. Space-resolved fluorescence microscopy was used to quantify PPIX fluorescence kinetics in the tumor and normal epithelium. Results: In cultures of normal epithelium, PPIX fluorescence kinetics were shown to depend on the duration of exposure to 5-ALA. There was a trend to higher fluorescence intensities with longer exposure times. In cocultures of bronchial epithelium and tumor, increases of fluorescence intensity were significantly greater in the tumor. Best tumor/normal tissue fluorescence ratios were found between 110 and 160 min after exposure to 5-ALA. Conclusion: Data obtained in this coculture system of bronchial epithelium and tumor is valuable to optimize modalities of fluorescence bronchoscopy for the diagnosis of early bronchial carcinoma.

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Secretory cell types and cell proliferation of human bronchial epithelial cells in an organ-culture system

Cited by 17 publications

References 27 publications

Polarized Expression of Cystic Fibrosis Transmembrane Conductance Regulator and Associated Epithelial Proteins during the Regeneration of Human Airway Surface Epithelium in Three-Dimensional Culture

Polarized Expression of Cystic Fibrosis Transmembrane Conductance Regulator and Associated Epithelial Proteins during the Regeneration of Human Airway Surface Epithelium in Three-Dimensional Culture

Repetitive exposure to zinc oxide nanoparticles induces dna damage in human nasal mucosa mini organ cultures

Kinetics of 5-Aminolevulinic Acid-Induced Fluorescence in Organ Cultures of Bronchial Epithelium and Tumor

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