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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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Time course of the impedance (a), acidification (b) and respiration (c) of L6 rat skeletal muscle cells during 24 h incubation with Acetaminophen in the Bionas 2500 Analyzing System. Vehicle control (black), 4.530 mg/L (30 mM, red), 1.510 mg/L (10 mM, blue), 755 mg/L (5 mM, green), 151 mg/L (1 mM, yellow) and 75.5 mg/L (0.5 mM, 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 AAP exposure (n = 3). (e) Results of BrdU assay of L6 cells incubated with AAP for 24 h.
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f6-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 Acetaminophen in the Bionas 2500 Analyzing System. Vehicle control (black), 4.530 mg/L (30 mM, red), 1.510 mg/L (10 mM, blue), 755 mg/L (5 mM, green), 151 mg/L (1 mM, yellow) and 75.5 mg/L (0.5 mM, 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 AAP exposure (n = 3). (e) Results of BrdU assay of L6 cells incubated with AAP for 24 h.

Mentions: AAP has an immediate effect on cellular respiration (see Figure 6(c)). Within 1 h after addition, AAP leads to a decrease of the respiration rate down to 30% for a concentration of 30 mM. Concentrations lower than 10 mM AAP show a fast recovery of the respiration (within 1 h) after 15 h of incubation. In contrast to the respiration, which shows a decrease for all AAP concentrations, the acidification shows a more complex signal pattern (see Figure 6(b)). Incubation with AAP concentrations lower than 5 mM results in a slight signal decrease compared to the control measurement. Higher concentrations (5–10 mM) activate the cell metabolism and lead to a strong increase of the acidification rates. The reduced capabilities of cells treated with 5–10 mM AAP to generate ATP via oxygen demanding oxidative phosphorylation, forces the cell to produce energy via increased glycolysis [46]. On the other hand, elevated levels of AAP (30 mM) lead also to a reduction of acidification. Excessive AAP toxification harms the cells permanently and therefore reduces all types of physiological activity. The impedance signals are constant over the whole tested concentration range (see Figure 6(a)). Concentrations of 30 mM lead to a decrease of 10–20% which is in good correlation with the reduced signals of the physiological parameters. Figure 6(d), showing the endpoint values of the changes relative to the control after 15 h of incubation with the various AAP concentrations, illustrates the opposing behavior of respiration and acidification at concentrations up to 10 mM.


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 Acetaminophen in the Bionas 2500 Analyzing System. Vehicle control (black), 4.530 mg/L (30 mM, red), 1.510 mg/L (10 mM, blue), 755 mg/L (5 mM, green), 151 mg/L (1 mM, yellow) and 75.5 mg/L (0.5 mM, 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 AAP exposure (n = 3). (e) Results of BrdU assay of L6 cells incubated with AAP for 24 h.
© Copyright Policy
Related In: Results  -  Collection

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

f6-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 Acetaminophen in the Bionas 2500 Analyzing System. Vehicle control (black), 4.530 mg/L (30 mM, red), 1.510 mg/L (10 mM, blue), 755 mg/L (5 mM, green), 151 mg/L (1 mM, yellow) and 75.5 mg/L (0.5 mM, 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 AAP exposure (n = 3). (e) Results of BrdU assay of L6 cells incubated with AAP for 24 h.
Mentions: AAP has an immediate effect on cellular respiration (see Figure 6(c)). Within 1 h after addition, AAP leads to a decrease of the respiration rate down to 30% for a concentration of 30 mM. Concentrations lower than 10 mM AAP show a fast recovery of the respiration (within 1 h) after 15 h of incubation. In contrast to the respiration, which shows a decrease for all AAP concentrations, the acidification shows a more complex signal pattern (see Figure 6(b)). Incubation with AAP concentrations lower than 5 mM results in a slight signal decrease compared to the control measurement. Higher concentrations (5–10 mM) activate the cell metabolism and lead to a strong increase of the acidification rates. The reduced capabilities of cells treated with 5–10 mM AAP to generate ATP via oxygen demanding oxidative phosphorylation, forces the cell to produce energy via increased glycolysis [46]. On the other hand, elevated levels of AAP (30 mM) lead also to a reduction of acidification. Excessive AAP toxification harms the cells permanently and therefore reduces all types of physiological activity. The impedance signals are constant over the whole tested concentration range (see Figure 6(a)). Concentrations of 30 mM lead to a decrease of 10–20% which is in good correlation with the reduced signals of the physiological parameters. Figure 6(d), showing the endpoint values of the changes relative to the control after 15 h of incubation with the various AAP concentrations, illustrates the opposing behavior of respiration and acidification at concentrations up to 10 mM.

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