Primary cultures of tracheal epithelial cells for the evaluation of respiratory toxicity

Blanquart, Catherine; Romet, Sylvie; Baeza, Armelle; Guennou, Christiane; Marano, Francelyne

doi:10.1016/0887-2333(91)90080-w

Cited by 17 publications

(9 citation statements)

References 11 publications

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“…Viability measurements (M'I'I" assay, neutral red uptake assay and release of cellular lactate dehydrogenase (LDH) assay) were carried out after 24 h of exposure to MTX, as described by Blanquart et al [8]. They were performed on 7-day-old cultures.…”

Section: Cell Viability Testsmentioning

confidence: 99%

Effects of maitotoxin on calcium entry and phosphoinositidebreakdown in the rabbit ciliated tracheal epithelium

Venant¹,

Dazy

Diogène

et al. 1994

Biology of the Cell

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Maitotoxin induces a concentration-dependent 45Ca uptake in primary cultures of rabbit tracheal epithelial cells. This response is insensitive to the calcium channel antagonists nifedipine, diltiazem and verapamil up to 20 microM. However, verapamil at 200 microM completely prevents 45Ca uptake. Measurements of indo-1 fluorescence show that MTX induces a very sustained (> or = 2 h) [Ca]i rise, which is completely inhibited by 200 microM of verapamil. Genistein (110 microM) (an inhibitor of tyrosine kinases) also strongly inhibits it. The inhibitory effect of 50 microM miconazole (an inhibitor of cytochrome P450) is only partial. Okadaic acid (inhibitor of protein-phosphatases) primarily delays the response to the toxin without decreasing its magnitude. MTX induces the formation of (1,4,5) inositol trisphosphate (IP3). The MTX response curve is biphasic. Stimulation is transient (< or = 10 min) and is not inhibited by chelation of intracellular Cai with BAPTA, nor by verapamil (200 microM) or U73122 (10 microM) (an inhibitor of activation of PLC beta 1 through a trimeric G protein). Results suggest that MTX independently activates a calcium transport process (which might imply phosphorylation on tyrosine residues) and a PLC not linked to a trimeric G protein.

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Section: Cell Viability Testsmentioning

confidence: 99%

Effects of maitotoxin on calcium entry and phosphoinositidebreakdown in the rabbit ciliated tracheal epithelium

Venant¹,

Dazy

Diogène

et al. 1994

Biology of the Cell

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“…We have shown that it is therefore possible to carry out a doseeffect study without difficulty for one or several molecules on the same trachea (Blanquart et al, 1991). On this model, we have improved a group of tests easy enough to carry out and which allows molecules to be classed according to their impact on the mucociliary epithelium.…”

Section: Discussionmentioning

confidence: 99%

“…MTT assay, neutral red uptake and release of cellular lactate dehydrogenase were performed as previously described (Blanquart et al, 1991). All these assays were made on days 7 and 8 on cells no longer proliferating (Baeza-Squiban et al, 1991).…”

Section: Viability Assaysmentioning

confidence: 99%

“…It enables the expansion of cell mass available for study and the rapid reconstitution of mucociliary epithelium maintaining the two characteristic differentiated cell types: secretory and ciliated cells. This in vitro system permits us to study cellular functions of normal respiratory epithelial cells and to investigate disturbances of those functions by a variety of pollutants (Romet et al, 1990;Baeza-Squiban et al, 1991;Blanquart et al, 1991).We report here on the development of in vitro mucociliary epithelial cell assays as methods of evaluating the specific toxicity of air pollutants.…”

Section: Introductionmentioning

confidence: 99%

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Tracheal epithelium in culture: a model for toxicity testing of inhaled molecules

et al. 1992

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Rabbit trachea primary cultures have been developed as a model to evaluate the toxicity of noxious airborne pollutants. A mucociliary epithelium has been restored in vitro on collagen gel. Several general cytotoxicity assays (viability and growth inhibition) permit a first assessment for the acute toxicity of the tested molecules. More specific criteria such as measurement of the integrity of the epithelial barrier and inhibition of ciliary beat frequency allow to determine a specific impact of xenobiotics on the mucociliary epithelium in culture.

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“…They include lung fibroblasts, tracheal cells (Mossman et al, 1983;Blanquart et al, 1991), pulmonary epithelial cells, and mesothelial cells (Jaurand et al, 1981). Mesothelial cells are of special interest because of the relationship between asbestos exposure and the occurrence of MM (Kamp et al, 1989) and of the absence of relationship with smoking.…”

Section: The Mesothelial Cell Modelmentioning

confidence: 99%

Use of mesothelial cell cultures to assess the carcinogenic potency of mineral or man made fibers

et al. 1992

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Natural mineral fibers may produce pulmonary cancers and mesothelioma. In contrast with lung cancer, the incidence of fiber-induced mesothelioma is not enhanced in smokers compared to non smokers. It is therefore of special interest to use mesothelial cells to study the toxicity of natural or man made mineral fibers. Several years ago, we have developed a method to culture rat pleural mesothelial cells (RPMC). We have first studied the effects of asbestos fibers by the application of in vitro tests formerly developed to determine the genotoxicity and transforming potency of soluble xenobiotics. Moreover, we have determined whether RPMC expressed cytochromes P450 known to metabolize polycyclic aromatic hydrocarbons. This paper reviews the results obtained so far. It has been found that asbestos fibers produced a cell transformation and a genotoxicity characterized by the formation of aneuploid cells, abnormal anaphases, chromosomal aberrations and DNA repair (UDS). In addition, RPMC expressed different forms of cytochromes P450. It is nowadays suggested that the tumorigenic potency of asbestos fibers may be related to the fiber dimensions, to their surface properties and in vivo biopersistence; this term involves the fiber solubility in biological medium and the fiber epuration from the lung by clearance mechanisms. Experiments are now in progress to determine whether the in vitro effects are dependent on the fiber parameters suggested as playing a role in the carcinogenic potency.

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Primary cultures of tracheal epithelial cells for the evaluation of respiratory toxicity

Cited by 17 publications

References 11 publications

Effects of maitotoxin on calcium entry and phosphoinositidebreakdown in the rabbit ciliated tracheal epithelium

Effects of maitotoxin on calcium entry and phosphoinositidebreakdown in the rabbit ciliated tracheal epithelium

Tracheal epithelium in culture: a model for toxicity testing of inhaled molecules

Use of mesothelial cell cultures to assess the carcinogenic potency of mineral or man made fibers

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