Eukaryotic cell lines as a sensitive layer for direct monitoring of carbon monoxide

Abstract

In the past years, whole cell gas toxicity assays have been developed in order to study the harmful effects of gaseous chemicals. Cells are incubated with nutrition medium supplemented with the gaseous substances to be tested. These methods are not feasible for the examination of toxic gases, which are insoluble in water. Carbon monoxide (CO), a poorly soluble and toxic gas (lethal dose 1 vol% for some minutes) has been chosen as a model gas. Besides its binding to hemoglobin, it also leads to an inhibition of cytochrome c oxidase in the mitochondrial respiratory chain of living cells. Thus, a decrease in the respiratory behavior of eukaryotic cells indicates the presence of CO in a gaseous phase. Based on the Bionas® 2500 analyzing system, a device to measure the metabolic and morphological effects, caused by direct gas exposure on eukaryotic cells was established. The gaseous substances directly contact the eukaryotic cell line V79 (Chinese hamster lung fibroblast) without interfering liquid phase in between. Beside the measurement of oxygen consumption also acidification and impedance changes of the cell culture are detected by a metabolic sensor chip. In order to increase the stability different chip surface coatings like collagen A and poly‐L‐lysin (PLL) were used. Adhesion coating leads to a tighter junction of the cells to the sensor chip surface and thus enables higher stability of the confluent cell monolayer towards the gaseous flow during the gas exposure. The method reported here has the potential to become a valuable means for rapid monitoring of toxic compounds in gaseous phases.

Using a cell-based gas biosensor for investigation of adverse effects of acetone vapors in vitro

“…The gas exposure measurement procedure differs from the measurements of solved acetone and were performed as described previously in detail (Bohrn et al 2011a). The metabolic sensor chips, covered with a confluent cell monolayer and 200 ml RM, are inserted in the biomodules of the 2500 analyzing system and connected to the reference electrode and the pumping system via the perfusion head.…”

“…1) which guides the gas stream over the cells to get it in contact with the cellular membrane via diffusion. Former studies (Bohrn et al 2010(Bohrn et al , 2011a demonstrated that exposure durations of 10-15 min are sufficient to generate detectable changes in the cellular signals, while the cells still rest on the sensor surface without being mechanically detached. For all experiments, reference measurements have to be done to compare the effects of the acetone gas with the influence of non-irritant humidified synthetic air.…”

A critical comparison of cell-based sensor systems for the detection of Cr(VI) in aquatic environment

“…The use of cell-based sensors has increased in the last few years [17]. Important areas of research are the toxicity monitoring in aquatic environments [18,19] and also the monitoring of toxic gaseous compounds in the air, which came into the focus of cell-based sensor research [20]. One of the main reasons for the recent success is the development of cellbased sensor systems which can be used in field applications [21].…”

Monitoring of irritant gas using a whole-cell-based sensor system

“…Until now, this system was primarily used for the detection of toxic substances in liquid medium. For this work, we adapted the system as reported earlier [40], so that it can be also used for the direct exposition of the mammalian cells with toxic substances in gas phases. A direct air-cell contact was generated by drainage of the liquid medium, thereby eliminating the possibility that gas molecules will be altered before contacting the cells [41].…”