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Cell-based sensor system using L6 cells for broad band continuous pollutant monitoring in aquatic environments.

Kubisch R, Bohrn U, Fleischer M, Stütz E - Sensors (Basel) (2012)

Bottom Line: In this study, the applicability of a cell-based sensor system using selected eukaryotic cell lines for the detection of aquatic pollutants is shown.A variety of potential cytotoxic classes of substances (heavy metals, pharmaceuticals, neurotoxins, waste water) was tested with monolayers of L6 cells (rat myoblasts).In a close to application model a real waste water sample shows detectable signals, indicating the existence of cytotoxic substances.

View Article: PubMed Central - PubMed

Affiliation: Pharmaceutical Biology-Biotechnology, Department of Pharmacy, Center for Drug Research, Ludwig-Maximilian-University Munich, Munich, Germany. rebekka.kubisch@cup.uni-muenchen.de

ABSTRACT
Pollution of drinking water sources represents a continuously emerging problem in global environmental protection. Novel techniques for real-time monitoring of water quality, capable of the detection of unanticipated toxic and bioactive substances, are urgently needed. In this study, the applicability of a cell-based sensor system using selected eukaryotic cell lines for the detection of aquatic pollutants is shown. Readout parameters of the cells were the acidification (metabolism), oxygen consumption (respiration) and impedance (morphology) of the cells. A variety of potential cytotoxic classes of substances (heavy metals, pharmaceuticals, neurotoxins, waste water) was tested with monolayers of L6 cells (rat myoblasts). The cytotoxicity or cellular effects induced by inorganic ions (Ni(2+) and Cu(2+)) can be detected with the metabolic parameters acidification and respiration down to 0.5 mg/L, whereas the detection limit for other substances like nicotine and acetaminophen are rather high, in the range of 0.1 mg/L and 100 mg/L. In a close to application model a real waste water sample shows detectable signals, indicating the existence of cytotoxic substances. The results support the paradigm change from single substance detection to the monitoring of overall toxicity.

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Related in: MedlinePlus

Time course of the impedance (a), acidification (b) and respiration (c) of L6 rat skeletal muscle cells during 24 h incubation with CuSO4 in the Bionas 2500 Analyzing System. Vehicle control (black), 10 mg/L (red), 5 mg/L (blue), 1 mg/L (green), 0.5 mg/L (yellow) and 0.1 mg/L (grey). Phases with running medium (RM) represent culture medium treatment without added substances. (d) Percent of control of impedance, respiration and acidification endpoint values measured after 15 h of copper sulfate exposure (n = 3). (e) Results of BrdU assay of L6 cells incubated with copper sulfate for 24 h.
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f5-sensors-12-03370: Time course of the impedance (a), acidification (b) and respiration (c) of L6 rat skeletal muscle cells during 24 h incubation with CuSO4 in the Bionas 2500 Analyzing System. Vehicle control (black), 10 mg/L (red), 5 mg/L (blue), 1 mg/L (green), 0.5 mg/L (yellow) and 0.1 mg/L (grey). Phases with running medium (RM) represent culture medium treatment without added substances. (d) Percent of control of impedance, respiration and acidification endpoint values measured after 15 h of copper sulfate exposure (n = 3). (e) Results of BrdU assay of L6 cells incubated with copper sulfate for 24 h.

Mentions: In the Bionas analyzing system, copper sulfate shows nearly no effect on the impedance signals of the L6 cells (see Figure 5(a)). In contrast the cellular metabolism is heavily altered by the copper ions which results in elevated acidification rates only at the beginning of the incubation at a concentration of 10 mg/L. Also other concentrations lower than 10 mg/L show a small increase in the acidification, compared to the untreated control measurements. Interestingly, the acidification rates decrease after this first increase (3 h after copper addition). The exposure to copper might lead to a first activation of the metabolism which is followed by another cellular response, tuning down the glycolysis (see Figure 5(b)). The cellular respiration rate is decreased at concentrations larger than 0.5 mg/L (see Figure 5(c)) which is supposedly caused by the oxidative stress due to reactive radicals. The respiration rates are able to recover at least partially after removal of copper sulfate containing media at the end of the experiment. Low concentrations of about 0.1 mg/L lead to a slight increase (Figure 5(d)).


Cell-based sensor system using L6 cells for broad band continuous pollutant monitoring in aquatic environments.

Kubisch R, Bohrn U, Fleischer M, Stütz E - Sensors (Basel) (2012)

Time course of the impedance (a), acidification (b) and respiration (c) of L6 rat skeletal muscle cells during 24 h incubation with CuSO4 in the Bionas 2500 Analyzing System. Vehicle control (black), 10 mg/L (red), 5 mg/L (blue), 1 mg/L (green), 0.5 mg/L (yellow) and 0.1 mg/L (grey). Phases with running medium (RM) represent culture medium treatment without added substances. (d) Percent of control of impedance, respiration and acidification endpoint values measured after 15 h of copper sulfate exposure (n = 3). (e) Results of BrdU assay of L6 cells incubated with copper sulfate for 24 h.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3376625&req=5

f5-sensors-12-03370: Time course of the impedance (a), acidification (b) and respiration (c) of L6 rat skeletal muscle cells during 24 h incubation with CuSO4 in the Bionas 2500 Analyzing System. Vehicle control (black), 10 mg/L (red), 5 mg/L (blue), 1 mg/L (green), 0.5 mg/L (yellow) and 0.1 mg/L (grey). Phases with running medium (RM) represent culture medium treatment without added substances. (d) Percent of control of impedance, respiration and acidification endpoint values measured after 15 h of copper sulfate exposure (n = 3). (e) Results of BrdU assay of L6 cells incubated with copper sulfate for 24 h.
Mentions: In the Bionas analyzing system, copper sulfate shows nearly no effect on the impedance signals of the L6 cells (see Figure 5(a)). In contrast the cellular metabolism is heavily altered by the copper ions which results in elevated acidification rates only at the beginning of the incubation at a concentration of 10 mg/L. Also other concentrations lower than 10 mg/L show a small increase in the acidification, compared to the untreated control measurements. Interestingly, the acidification rates decrease after this first increase (3 h after copper addition). The exposure to copper might lead to a first activation of the metabolism which is followed by another cellular response, tuning down the glycolysis (see Figure 5(b)). The cellular respiration rate is decreased at concentrations larger than 0.5 mg/L (see Figure 5(c)) which is supposedly caused by the oxidative stress due to reactive radicals. The respiration rates are able to recover at least partially after removal of copper sulfate containing media at the end of the experiment. Low concentrations of about 0.1 mg/L lead to a slight increase (Figure 5(d)).

Bottom Line: In this study, the applicability of a cell-based sensor system using selected eukaryotic cell lines for the detection of aquatic pollutants is shown.A variety of potential cytotoxic classes of substances (heavy metals, pharmaceuticals, neurotoxins, waste water) was tested with monolayers of L6 cells (rat myoblasts).In a close to application model a real waste water sample shows detectable signals, indicating the existence of cytotoxic substances.

View Article: PubMed Central - PubMed

Affiliation: Pharmaceutical Biology-Biotechnology, Department of Pharmacy, Center for Drug Research, Ludwig-Maximilian-University Munich, Munich, Germany. rebekka.kubisch@cup.uni-muenchen.de

ABSTRACT
Pollution of drinking water sources represents a continuously emerging problem in global environmental protection. Novel techniques for real-time monitoring of water quality, capable of the detection of unanticipated toxic and bioactive substances, are urgently needed. In this study, the applicability of a cell-based sensor system using selected eukaryotic cell lines for the detection of aquatic pollutants is shown. Readout parameters of the cells were the acidification (metabolism), oxygen consumption (respiration) and impedance (morphology) of the cells. A variety of potential cytotoxic classes of substances (heavy metals, pharmaceuticals, neurotoxins, waste water) was tested with monolayers of L6 cells (rat myoblasts). The cytotoxicity or cellular effects induced by inorganic ions (Ni(2+) and Cu(2+)) can be detected with the metabolic parameters acidification and respiration down to 0.5 mg/L, whereas the detection limit for other substances like nicotine and acetaminophen are rather high, in the range of 0.1 mg/L and 100 mg/L. In a close to application model a real waste water sample shows detectable signals, indicating the existence of cytotoxic substances. The results support the paradigm change from single substance detection to the monitoring of overall toxicity.

Show MeSH
Related in: MedlinePlus