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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

Example of RTCA profiles generated with etoposide (A549 and HepG2 cells) and proprietary UCB compounds (HepG2 cells). (A) HepG2 cells were exposed to: 0 (0.5% DMSO, red curve), 0.1 (green curve), 1 (purple curve), 10 (dark blue curve) and 100 (light blue curve) µM of etoposide. (B) A549 cells exposed to 0 (control DMSO, red curve) and 20 µM (green curve) of etoposide (Abassi et al. [9]). (C–F): HepG2 cells were exposed to 0 (0.5% DMSO, red curve), 125 (green curve), 250 (purple curve), 500 (dark blue curve) and 1,000 (light blue curve) µM of compound UCB 1, 2, 3 and 4, respectively. Cell indexes were normalized with the last time point before compound addition. The normalized time point is indicated by the vertical line. All panels (except B): each data point was calculated from triplicate values (except for control cells n = 6). Data represent the average ± standard deviation (except for panel B). A zoom is provided in case of overlapping curves. For more details, please refer to the Materials and Methods section.
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biosensors-03-00132-f007: Example of RTCA profiles generated with etoposide (A549 and HepG2 cells) and proprietary UCB compounds (HepG2 cells). (A) HepG2 cells were exposed to: 0 (0.5% DMSO, red curve), 0.1 (green curve), 1 (purple curve), 10 (dark blue curve) and 100 (light blue curve) µM of etoposide. (B) A549 cells exposed to 0 (control DMSO, red curve) and 20 µM (green curve) of etoposide (Abassi et al. [9]). (C–F): HepG2 cells were exposed to 0 (0.5% DMSO, red curve), 125 (green curve), 250 (purple curve), 500 (dark blue curve) and 1,000 (light blue curve) µM of compound UCB 1, 2, 3 and 4, respectively. Cell indexes were normalized with the last time point before compound addition. The normalized time point is indicated by the vertical line. All panels (except B): each data point was calculated from triplicate values (except for control cells n = 6). Data represent the average ± standard deviation (except for panel B). A zoom is provided in case of overlapping curves. For more details, please refer to the Materials and Methods section.

Mentions: Evaluation of RTCA predictivity to detect genotoxicity with a set of 81 proprietary UCB compounds. HepG2 cells were exposed for at least 72 h to 81 proprietary UCB compounds belonging to two central nervous system projects (CNS1 and CNS2). The concentrations tested were 125, 250, 500 and 1,000 µM (unless solubility problems were encountered). The objective was to determine if the compounds that were identified as genotoxic by traditional genotoxicity in vitro assays generated RTCA genotoxic profiles. A compound was classified as genotoxic according to the impedance measurements when after the compound addition, the cell index generated by the compound was higher than the control curve followed by a decrease in cell index that reached at least 50% mortality within 48 h exposure. A compound was classified as negative when both conditions were not met. Examples of typical genotoxicity profiles are presented in Figure 7(A,B) (i.e., HepG2 and A549 cells exposed to etoposide, respectively). For more details about the definition of the terms (sensitivity, specificity and concordance/accuracy), please refer to the Materials and Methods section.


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)

Example of RTCA profiles generated with etoposide (A549 and HepG2 cells) and proprietary UCB compounds (HepG2 cells). (A) HepG2 cells were exposed to: 0 (0.5% DMSO, red curve), 0.1 (green curve), 1 (purple curve), 10 (dark blue curve) and 100 (light blue curve) µM of etoposide. (B) A549 cells exposed to 0 (control DMSO, red curve) and 20 µM (green curve) of etoposide (Abassi et al. [9]). (C–F): HepG2 cells were exposed to 0 (0.5% DMSO, red curve), 125 (green curve), 250 (purple curve), 500 (dark blue curve) and 1,000 (light blue curve) µM of compound UCB 1, 2, 3 and 4, respectively. Cell indexes were normalized with the last time point before compound addition. The normalized time point is indicated by the vertical line. All panels (except B): each data point was calculated from triplicate values (except for control cells n = 6). Data represent the average ± standard deviation (except for panel B). A zoom is provided in case of overlapping curves. 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-f007: Example of RTCA profiles generated with etoposide (A549 and HepG2 cells) and proprietary UCB compounds (HepG2 cells). (A) HepG2 cells were exposed to: 0 (0.5% DMSO, red curve), 0.1 (green curve), 1 (purple curve), 10 (dark blue curve) and 100 (light blue curve) µM of etoposide. (B) A549 cells exposed to 0 (control DMSO, red curve) and 20 µM (green curve) of etoposide (Abassi et al. [9]). (C–F): HepG2 cells were exposed to 0 (0.5% DMSO, red curve), 125 (green curve), 250 (purple curve), 500 (dark blue curve) and 1,000 (light blue curve) µM of compound UCB 1, 2, 3 and 4, respectively. Cell indexes were normalized with the last time point before compound addition. The normalized time point is indicated by the vertical line. All panels (except B): each data point was calculated from triplicate values (except for control cells n = 6). Data represent the average ± standard deviation (except for panel B). A zoom is provided in case of overlapping curves. For more details, please refer to the Materials and Methods section.
Mentions: Evaluation of RTCA predictivity to detect genotoxicity with a set of 81 proprietary UCB compounds. HepG2 cells were exposed for at least 72 h to 81 proprietary UCB compounds belonging to two central nervous system projects (CNS1 and CNS2). The concentrations tested were 125, 250, 500 and 1,000 µM (unless solubility problems were encountered). The objective was to determine if the compounds that were identified as genotoxic by traditional genotoxicity in vitro assays generated RTCA genotoxic profiles. A compound was classified as genotoxic according to the impedance measurements when after the compound addition, the cell index generated by the compound was higher than the control curve followed by a decrease in cell index that reached at least 50% mortality within 48 h exposure. A compound was classified as negative when both conditions were not met. Examples of typical genotoxicity profiles are presented in Figure 7(A,B) (i.e., HepG2 and A549 cells exposed to etoposide, respectively). For more details about the definition of the terms (sensitivity, specificity and concordance/accuracy), please refer to the Materials and Methods section.

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