Minireview: Culture of Airway Epithelial Cells: Research Techniques

Scott, Michael R. Van; Yankaskas, James R.; Boucher, Richard C.

doi:10.3109/01902148609063272

Cited by 51 publications

(13 citation statements)

References 86 publications

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“…Gene transfer to primary cultures of cotton rat airway epithelial cells Cultures of cotton rat airway epithelial cells were prepared by described methods (Van Scott et al, 1986). Dissociated cells were harvested, washed three times with F12-7X media, and plated at a density of 5.0 x 10^ cells/dish in 3-cm tissue culture dishes.…”

Section: Introductionmentioning

confidence: 99%

DirectIn VivoGene Transfer to Airway Epithelium Employing Adenovirus–Polylysine–DNA Complexes

Gao¹,

Wagner²,

Cotten³

et al. 1993

Human Gene Therapy

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Adenovirus-polylysine-DNA complexes were evaluated for their capacity to accomplish direct in vivo gene transfer to airway epithelium employing a rodent model. Binary complexes containing transferrin or adenovirus, or combination complexes containing both transferrin and adenovirus, were evaluated. The highest in vitro gene transfer efficiency in primary cultures of airway epithelial cells was accomplished by the combination complexes. This result was paralleled in vivo. Transient gene expression of up to 1 week was observed with localization of the transduced cells to the region of the small airways. These results establish the feasibility of this type of approach for gene therapy applications.

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Section: Introductionmentioning

confidence: 99%

DirectIn VivoGene Transfer to Airway Epithelium Employing Adenovirus–Polylysine–DNA Complexes

Gao¹,

Wagner²,

Cotten³

et al. 1993

Human Gene Therapy

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“…Of the many tracheal and bronchial epithelial culture systems which have been developed, a classical system culturing TBECs on plastic dishes without coating by collagen gels or extracellular matrix was used in this Chemokine expression by cultured epithelial cells 89 study. Preservation of the morphological differentiation of the cells grown on permeable supports or the cells repopulated with heterologous grafts is apparently better than with cells grown on nontreated dishes, and airway epithelial cells grown on nontreated dishes are generally nonciliated and squamoid; however, the cells grown on nontreated dishes do maintain many functional characteristics (Van Scott et al 1986). Since we aimed to evaluate the function of morphologically altered cells such as the squamoid cells in vitro, we tried out to utilize this culture system.…”

Section: Discussionmentioning

confidence: 99%

Morphological Alteration of Cultured Tracheobronchial Epithelial Cells is Accompanied by the Expression of Chemokines, MCP‐1 and CINC/gro, in Rats

Yamamoto

Iyonaga²,

Takeya³

et al. 1998

Int J Experimental Path

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Morphologically altered epithelial cells are generally observed in fibrotic lung conditions and have been reported to produce several cytokines. To examine the relationship between morphological changes of the tracheobronchial epithelial cells (TBECs) and their chemokine production, we investigated, (1) the mRNA expression and protein secretion of monocyte chemoattractant protein-1 (MCP-1) and cytokine-induced neutrophil chemoattractant/gro (CINC/gro), (2) morphological changes by electron microscopy, and (3) cytokeratin (CK) expression, using a primary culture system of rat TBECs. The constitutive secretion of MCP-1 in the culture supernatant of TBECs increased in a time-dependent manner, whereas the CINC/gro secretion was not changed. These results were consistent with the chemokines' mRNA expression observed by in situ hybridization. The constitutive secretions of MCP-1 and CINC/gro were inhibited partially but significantly by dexamethasone. With the extension of the culture period, the morphology of the TBECs became flat and spindle in shape, similar to squamous metaplasia, as observed on electron microscopy, and with strong expression of CK 14. Sequential staining using immunocytochemistry and in situ hybridization revealed the coexpression of MCP-1 mRNA and CK 14. These data indicate a significant relationship between the morphological squamoid alteration and the constitutive expression of MCP-1 but not of CINC/gro. It is thought that the squamous metaplasia of TBECs may accompany the alteration of cytokine production and play an important role in chronic lung inflammation.

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“…In epithelial cells, the expression of complete differentiation requires the development of polarity within the cell (Sattler et al, 1978;Chambard et al, 1982), although to what extent polarity is achievable in transformed cells is as yet unclear. It has been reported that for the correct expression of differentiated features in normal lung, it is often necessary to have the cells growing close to the air-liquid interface, since this mimics more closely the in vivo situation (Van Scott et al, 1986), so elevating the culture closer to the surface of the medium may have contributed to increased PS synthesis.…”

Section: Discussionmentioning

confidence: 99%

Paracrine control of differentiation in the alveolar carcinoma, A549, by human foetal lung fibroblasts

Speirs

Ray²,

Freshney³

1991

Br J Cancer

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Summary Synthesis of pulmonary surfactant (PS) is necessary for normal functioning of the lungs and its production is indicative of normal differentiated lung. The human alveolar carcinoma, A549, has been found to synthesis and secrete PS in vitro. The purpose of this study was to optimise the culture conditions for PS synthesis by A549 as well as to determine the potential role of foetal lung fibroblasts in the induction of PS by glucocorticoids. A549 cells growing in filter wells produced higher levels of PS in response to steroid, a 5-fold increase on the filter well compared to only a 1.5-fold increase when the cells were cultured on a conventional plastic substrate. A549 cells grown in filter wells responded to coculture with fibroblasts whether in direct contact or separated co-culture. A 20-fold increase in PS over control values was observed in separated steroid-treated co-cultures, suggesting the presence of a diffusible factor. A partially purified factor was isolated from fibroblast conditioned medium which was capable of inducing differentiation and other phenotypic changes in A549, namely induction of PS, reduction of plasminogen activator activity and reduction in the in vivo growth of A549 xenografts in nude mice. These results suggest that, under the correct conditions, A549 cells, although transformed, still retain the capacity to respond to differentiation-inducing signals from normal fibroblasts.

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Minireview: Culture of Airway Epithelial Cells: Research Techniques

Cited by 51 publications

References 86 publications

DirectIn VivoGene Transfer to Airway Epithelium Employing Adenovirus–Polylysine–DNA Complexes

DirectIn VivoGene Transfer to Airway Epithelium Employing Adenovirus–Polylysine–DNA Complexes

Morphological Alteration of Cultured Tracheobronchial Epithelial Cells is Accompanied by the Expression of Chemokines, MCP‐1 and CINC/gro, in Rats

Paracrine control of differentiation in the alveolar carcinoma, A549, by human foetal lung fibroblasts

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