Online monitoring of cell metabolism for studying pharmacodynamic effects

Thedinga, Elke; Kob, A.; Holst, Heiko; Keuer, Andreas; Drechsler, S.; Niendorf, Ricarda; Baumann, Werner; Freund, Ingo; Lehmann, Mirko; Ehret, Ralf

doi:10.1016/j.taap.2006.12.027

Cited by 74 publications

(60 citation statements)

References 11 publications

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“…Interestingly, also NK11, the inactive variant of NK-2, led to a similar increase in impedance upon addition to LA-N-1 cells. Such an increase in impedance has been observed rarely and it has been shown that it is not associated with cell damage, or an effect on the organization of the cytoskeleton (Ceriotti et al 2007;Thedinga et al 2007), which led to a decrease in impedance, but with cycloheximide-mediated inhibition of protein biosynthesis in HT-29 cells ), suggesting intracellular structures as secondary targets for NK-2. Among the three peptides, melittin was most effective.…”

Section: Discussionmentioning

confidence: 99%

“…Since they are normally evaluated after a certain time point (endpoint), they will overlook kinetics as well as cell damages, which occur at an early time point and which are repaired by the cells during incubation time. To close this gap in the understanding of the mode of action of AMPs as anti-cancer agents, we utilized a chip-based analyzing system, which allows the continuous and label free determination of cell metabolism and cell morphology over time (Abarzua et al 2010;Ceriotti et al 2007;Thedinga et al 2007). Cells were grown on a chip surface, which is equipped with pH, oxygen and impedance sensors.…”

Section: Introductionmentioning

confidence: 99%

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Online monitoring of metabolism and morphology of peptide-treated neuroblastoma cancer cells and keratinocytes

Drechsler¹,

Andrä

2011

J Bioenerg Biomembr

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Antimicrobial peptides are promising anti-cancer agents with a unique mode of action. We established the usage of a chip-based sensor to monitor the dynamic interplay between cells on the chip and peptides and compared it with endpoint tests. Human neuroblastoma cancer cells and spontaneously immortalized non-cancer keratinocytes were perfused with representative peptides (NK-2, NK11, and melittin). The sensor system enabled continuous recording of cell layer impedance (adhesion/ confluence), oxygen consumption (respiration) and extracellular acidification (glycolysis) and provided insights in cell damage, stress response and recovery. Cells responded differentially to peptide treatment. During perfusion, peptides accumulated on the cell surface until they reached a critical concentration. Preceding to cell death, melittin triggered glycolysis, suggesting stress response. NK-2 induced no change in energy metabolism, but led to an increase in impedance, i.e. a temporarily altered morphology, which appeared to be an excellent parameter to detect subtle structural changes of cell layers.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Online monitoring of metabolism and morphology of peptide-treated neuroblastoma cancer cells and keratinocytes

Drechsler¹,

Andrä

2011

J Bioenerg Biomembr

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“…The metabolic activity of cells can be monitored by measuring the acidification rate in the extracellular environment; therefore, changes in the biological function and the response to ligand binding to surface receptors, and the impact of chemicals and drugs on cell physiology can be studied by measuring the decrease in pH that occurs as acidic metabolites build up in the extracellular space (Lorenzelli et al, 2003;Parak et al, 2000;Thedinga et al, 2007). volumes using a Light Addressable Potentiometric Sensor to determine acidification rates in order to assess the metabolic activity of cells during pharma-or toxicological studies (Eklund et al, 2003;Hafner, 2000;Owicki et al, 1994).…”

Section: Introductionmentioning

confidence: 99%

On-chip acidification rate measurements from single cardiac cells confined in sub-nanoliter volumes

Ges

Dzhura

Baudenbacher

2008

Biomed Microdevices

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The metabolic activity of cells can be monitored by measuring the pH in the extracellular environment. Microfabrication and microfluidic technologies allow the sensor size and the extracellular volumes to be comparable to single cells. A glass substrate with thin film pH sensitive IrO x electrodes was sealed to a replica-molded polydimethylsiloxane (PDMS) microfluidic network with integrated valves. The device, termed NanoPhysiometer, allows the trapping of single cardiac myocytes and the measurement of the pH in a detection volume of 0.36 nL. For wild-type (WT) single cardiac myocytes an acidification rate of 6.45 ± 0.38 mpH/min was measured in comparison to 19.5 ± 0.38 mpH/min for very long chain Acyl-CoA dehydrogenase (VLCAD) deficient mice in 0.8 mM of Ca 2+ . VLCAD deficiency is a fatty acid oxidation disease leading to cardiomyopathy and arrhythmias. The acidification rate increased to 11.96 ± 1.33 mpH/min for WT and to 32.0 ± 4.64 mpH/min for VLCAD −/− in 1.8 mM of Ca 2+ . The NanoPhysiometer concept can be extended to study ischemia/reperfusion injury or disorders of other biological systems to identify strategies for treatment and possible pharmacological targets.

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“…Live cell monitoring of the adhesion and acidification under the influence of compound 1 was performed with the Bionas Õ 2500 analyzing system with the metabolic chip SC 1000 (Bionas GmbH, Rostock, Germany) and the measurement software Bionas15002 CS1.47 (Thedinga et al, 2007). Prior to experiments, chips were cleaned with 70% ethanol for 10 min, washed with PBS and were adapted to the measurement medium for 5 min.…”

Section: Online Monitoring Of Metabolic Alterationsmentioning

confidence: 99%

“…On each chip (2 Â 10 6 ), cells were seeded and allowed to adhere overnight at 37 C and in 5% CO 2 so that 80% confluence on the sensor chips was reached. Bionas measurements were carried out with a pump rate of 56 mL/ min for 24 h (Rebl et al, 2010;Thedinga et al, 2007). Within the first 4 h, cells could adapt to the new measurement medium.…”

Section: Online Monitoring Of Metabolic Alterationsmentioning

confidence: 99%

Novel anticancer alkene lactone fromPersea americana

Falodun¹,

Engel

Kragl³

et al. 2013

Pharmaceutical Biology

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Context: Persea americana Mill (Lauraceae) root bark is used in ethnomedicine for a variety of diseases including cancer. Objective: To isolate and characterize the chemical constituent in P. americana, and also to determine the anticancer property of a new alkene lactone from the root bark of P. americana. Materials and methods: The MCF-7 cells were treated with different concentrations of the pure compound for 48 h. The percentage of cells in the various phases, online monitoring of metabolic changes and integrin receptor expression determined by flow cytometry. Results: One novel alkene lactone (4-hydroxy-5-methylene-3-undecyclidenedihydrofuran-2 (3H)-one) (1) was isolated and characterized using 1D-NMR, 2D-NMR, infrared, UV and MS. At a concentration of 10 mg/mL, significant reduction of proliferation of MCF-7 was induced while MCF-12 A cell was significantly stimulated by 10 mg/mL. The IC 50 value for MCF-7 cells is 20.48 mg/mL. Lower concentration of 1 harbor no significant effect on either MCF-7 or MCF-12A. The apoptotic rates of MCF-7 cells were increased significantly. At the final concentration 10 mg/mL, up to 80% of all breast cancer cells were dead. On the non-tumorigenic cell line MCF-12A, the same concentrations (1 and 10 mg/mL) of compound 1 caused significant enhanced apoptotic rates. A total of 1 mg/mL of 1 caused a decrease of a4-, a6-, b1-and b3-integrin expression. Conclusions: The compound caused a stimulatory effect on non-tumorigenic MCF-12A cells with respect to cell adhesion while tumorigenic MCF-7 cells detached continuously. This is the first report on the anticancer effects of this class of compound.

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Online monitoring of cell metabolism for studying pharmacodynamic effects

Cited by 74 publications

References 11 publications

Online monitoring of metabolism and morphology of peptide-treated neuroblastoma cancer cells and keratinocytes

Online monitoring of metabolism and morphology of peptide-treated neuroblastoma cancer cells and keratinocytes

On-chip acidification rate measurements from single cardiac cells confined in sub-nanoliter volumes

Novel anticancer alkene lactone fromPersea americana

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