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Evaluation of impedance-based label-free technology as a tool for pharmacology and toxicology investigations.

Atienzar FA, Gerets H, Tilmant K, Toussaint G, Dhalluin S - Biosensors (Basel) (2013)

Bottom Line: In addition, specific RTCA profiles (signatures) were generated when HepG2 and HepaRG cells were exposed to calcium modulators, antimitotics, DNA damaging and nuclear receptor agents, with a percentage of prediction close to 80% for both cellular models.In a subsequent experiment, HepG2 cells were exposed to 81 proprietary UCB compounds known to be genotoxic or not.Based on the DNA damaging signatures, the RTCA technology allowed the detection of ca. 50% of the genotoxic compounds (n = 29) and nearly 100% of the non-genotoxic compounds (n = 52).

View Article: PubMed Central - PubMed

Affiliation: UCB Pharma SA, Non Clinical Development, Chemin du Foriest, 1420 Braine-l'Alleud, Belgium; E-Mails: helga.gerets@ucb.com (H.G.); karen.tilmant@ucb.com (K.T.); gaelle.toussaint@ucb.com (G.T.); stephane.dhalluin@ucb.com (S.D.).

ABSTRACT
The use of label-free technologies based on electrical impedance is becoming more and more popular in drug discovery. Indeed, such a methodology allows the continuous monitoring of diverse cellular processes, including proliferation, migration, cytotoxicity and receptor-mediated signaling. The objective of the present study was to further assess the usefulness of the real-time cell analyzer (RTCA) and, in particular, the xCELLigence platform, in the context of early drug development for pharmacology and toxicology investigations. In the present manuscript, four cellular models were exposed to 50 compounds to compare the cell index generated by RTCA and cell viability measured with a traditional viability assay. The data revealed an acceptable correlation (ca. 80%) for both cell lines (i.e., HepG2 and HepaRG), but a lack of correlation (ca. 55%) for the primary human and rat hepatocytes. In addition, specific RTCA profiles (signatures) were generated when HepG2 and HepaRG cells were exposed to calcium modulators, antimitotics, DNA damaging and nuclear receptor agents, with a percentage of prediction close to 80% for both cellular models. In a subsequent experiment, HepG2 cells were exposed to 81 proprietary UCB compounds known to be genotoxic or not. Based on the DNA damaging signatures, the RTCA technology allowed the detection of ca. 50% of the genotoxic compounds (n = 29) and nearly 100% of the non-genotoxic compounds (n = 52). Overall, despite some limitations, the xCELLigence platform is a powerful and reliable tool that can be used in drug discovery for toxicity and pharmacology studies.

No MeSH data available.


Related in: MedlinePlus

RTCA profiles generated in HepG2, cryopreserved HepaRG, primary rat and human hepatocytes exposed to cerivastatin. Cellular models as indicated in the panels A, B, C and D. Cells were exposed to: 0 (0.5% DMSO, red curve), 33 (green curve), 66 (purple curve), 132 (dark blue curve) and 264 (light blue curve) µM cerivastatin for five days. The concentration tested were multiple of the Cmax value (Cmax cerivastatin = 2.64 µM), i.e., 12.5, 25, 50 and 100 Cmax. Cerivastatin was added on day 0 (close to the vertical line) and day two. Cell indexes were normalized with the last time point before compound addition. The normalized time point is indicated by the vertical line. Each data point was calculated from triplicate values (except for control cells n = 6). Data represent the average ± standard deviation. For more details, please refer to the Materials and Methods section.
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biosensors-03-00132-f002: RTCA profiles generated in HepG2, cryopreserved HepaRG, primary rat and human hepatocytes exposed to cerivastatin. Cellular models as indicated in the panels A, B, C and D. Cells were exposed to: 0 (0.5% DMSO, red curve), 33 (green curve), 66 (purple curve), 132 (dark blue curve) and 264 (light blue curve) µM cerivastatin for five days. The concentration tested were multiple of the Cmax value (Cmax cerivastatin = 2.64 µM), i.e., 12.5, 25, 50 and 100 Cmax. Cerivastatin was added on day 0 (close to the vertical line) and day two. Cell indexes were normalized with the last time point before compound addition. The normalized time point is indicated by the vertical line. Each data point was calculated from triplicate values (except for control cells n = 6). Data represent the average ± standard deviation. For more details, please refer to the Materials and Methods section.

Mentions: The RTCA curves show that the cell index of control cells exposed to 0.5% DMSO increased and/or stabilized for all cell types up to five days of treatment except for the primary rat hepatocytes (Figure 2). Indeed, for this latter model, the cell index started to decline particularly after two-days’ exposure (Figure 2(D)). Dose response relationships were obtained particularly for HepG2 cells (Figure 2(A)) and to a lesser degree for HepaRG cells (Figure 2(B)) exposed to cerivastatin. Significant cytotoxic effects were observed for all concentrations tested in primary human hepatocytes exposed to cerivastatin (Figure 2(C)). For all the cellular models tested, clear cytotoxic effects were already observed after the first exposure period at all cerivastatin concentrations tested. The addition of cerivastatin a second time on day two did not induce a more significant decrease in cell index (Figure 2) for the four cellular models.


Evaluation of impedance-based label-free technology as a tool for pharmacology and toxicology investigations.

Atienzar FA, Gerets H, Tilmant K, Toussaint G, Dhalluin S - Biosensors (Basel) (2013)

RTCA profiles generated in HepG2, cryopreserved HepaRG, primary rat and human hepatocytes exposed to cerivastatin. Cellular models as indicated in the panels A, B, C and D. Cells were exposed to: 0 (0.5% DMSO, red curve), 33 (green curve), 66 (purple curve), 132 (dark blue curve) and 264 (light blue curve) µM cerivastatin for five days. The concentration tested were multiple of the Cmax value (Cmax cerivastatin = 2.64 µM), i.e., 12.5, 25, 50 and 100 Cmax. Cerivastatin was added on day 0 (close to the vertical line) and day two. Cell indexes were normalized with the last time point before compound addition. The normalized time point is indicated by the vertical line. Each data point was calculated from triplicate values (except for control cells n = 6). Data represent the average ± standard deviation. For more details, please refer to the Materials and Methods section.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263586&req=5

biosensors-03-00132-f002: RTCA profiles generated in HepG2, cryopreserved HepaRG, primary rat and human hepatocytes exposed to cerivastatin. Cellular models as indicated in the panels A, B, C and D. Cells were exposed to: 0 (0.5% DMSO, red curve), 33 (green curve), 66 (purple curve), 132 (dark blue curve) and 264 (light blue curve) µM cerivastatin for five days. The concentration tested were multiple of the Cmax value (Cmax cerivastatin = 2.64 µM), i.e., 12.5, 25, 50 and 100 Cmax. Cerivastatin was added on day 0 (close to the vertical line) and day two. Cell indexes were normalized with the last time point before compound addition. The normalized time point is indicated by the vertical line. Each data point was calculated from triplicate values (except for control cells n = 6). Data represent the average ± standard deviation. For more details, please refer to the Materials and Methods section.
Mentions: The RTCA curves show that the cell index of control cells exposed to 0.5% DMSO increased and/or stabilized for all cell types up to five days of treatment except for the primary rat hepatocytes (Figure 2). Indeed, for this latter model, the cell index started to decline particularly after two-days’ exposure (Figure 2(D)). Dose response relationships were obtained particularly for HepG2 cells (Figure 2(A)) and to a lesser degree for HepaRG cells (Figure 2(B)) exposed to cerivastatin. Significant cytotoxic effects were observed for all concentrations tested in primary human hepatocytes exposed to cerivastatin (Figure 2(C)). For all the cellular models tested, clear cytotoxic effects were already observed after the first exposure period at all cerivastatin concentrations tested. The addition of cerivastatin a second time on day two did not induce a more significant decrease in cell index (Figure 2) for the four cellular models.

Bottom Line: In addition, specific RTCA profiles (signatures) were generated when HepG2 and HepaRG cells were exposed to calcium modulators, antimitotics, DNA damaging and nuclear receptor agents, with a percentage of prediction close to 80% for both cellular models.In a subsequent experiment, HepG2 cells were exposed to 81 proprietary UCB compounds known to be genotoxic or not.Based on the DNA damaging signatures, the RTCA technology allowed the detection of ca. 50% of the genotoxic compounds (n = 29) and nearly 100% of the non-genotoxic compounds (n = 52).

View Article: PubMed Central - PubMed

Affiliation: UCB Pharma SA, Non Clinical Development, Chemin du Foriest, 1420 Braine-l'Alleud, Belgium; E-Mails: helga.gerets@ucb.com (H.G.); karen.tilmant@ucb.com (K.T.); gaelle.toussaint@ucb.com (G.T.); stephane.dhalluin@ucb.com (S.D.).

ABSTRACT
The use of label-free technologies based on electrical impedance is becoming more and more popular in drug discovery. Indeed, such a methodology allows the continuous monitoring of diverse cellular processes, including proliferation, migration, cytotoxicity and receptor-mediated signaling. The objective of the present study was to further assess the usefulness of the real-time cell analyzer (RTCA) and, in particular, the xCELLigence platform, in the context of early drug development for pharmacology and toxicology investigations. In the present manuscript, four cellular models were exposed to 50 compounds to compare the cell index generated by RTCA and cell viability measured with a traditional viability assay. The data revealed an acceptable correlation (ca. 80%) for both cell lines (i.e., HepG2 and HepaRG), but a lack of correlation (ca. 55%) for the primary human and rat hepatocytes. In addition, specific RTCA profiles (signatures) were generated when HepG2 and HepaRG cells were exposed to calcium modulators, antimitotics, DNA damaging and nuclear receptor agents, with a percentage of prediction close to 80% for both cellular models. In a subsequent experiment, HepG2 cells were exposed to 81 proprietary UCB compounds known to be genotoxic or not. Based on the DNA damaging signatures, the RTCA technology allowed the detection of ca. 50% of the genotoxic compounds (n = 29) and nearly 100% of the non-genotoxic compounds (n = 52). Overall, despite some limitations, the xCELLigence platform is a powerful and reliable tool that can be used in drug discovery for toxicity and pharmacology studies.

No MeSH data available.


Related in: MedlinePlus