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Time-resolved cell culture assay analyser (TReCCA Analyser) for the analysis of on-line data: data integration--sensor correction--time-resolved IC50 determination.

Lochead J, Schessner J, Werner T, Wölfl S - PLoS ONE (2015)

Bottom Line: The functions of the program include data normalising and averaging, as well as smoothing and slope calculation, pin-pointing exact change time points.To illustrate the capabilities of the TReCCA Analyser, we performed on-line monitoring of dissolved oxygen in the culture media of the breast cancer cell line MCF-7 treated with different concentrations of the anti-cancer drug Cisplatin.The TReCCA Analyser is freely available at www.uni-heidelberg.de/fakultaeten/biowissenschaften/ipmb/biologie/woelfl/Research.html.

View Article: PubMed Central - PubMed

Affiliation: Institute for Pharmacy and Molecular Biotechnology, Heidelberg University, Heidelberg, Germany; Institute for Analytical Chemistry, Mannheim University of Applied Sciences, Mannheim, Germany.

ABSTRACT
Time-resolved cell culture assays circumvent the need to set arbitrary end-points and reveal the dynamics of quality controlled experiments. However, they lead to the generation of large data sets, which can represent a complexity barrier to their use. We therefore developed the Time-Resolved Cell Culture Assay (TReCCA) Analyser program to perform standard cell assay analyses efficiently and make sophisticated in-depth analyses easily available. The functions of the program include data normalising and averaging, as well as smoothing and slope calculation, pin-pointing exact change time points. A time-resolved IC50/EC50 calculation provides a better understanding of drug toxicity over time and a more accurate drug to drug comparison. Finally the logarithmic sensor recalibration function, for sensors with an exponential calibration curve, homogenises the sensor output and enables the detection of low-scale changes. To illustrate the capabilities of the TReCCA Analyser, we performed on-line monitoring of dissolved oxygen in the culture media of the breast cancer cell line MCF-7 treated with different concentrations of the anti-cancer drug Cisplatin. The TReCCA Analyser is freely available at www.uni-heidelberg.de/fakultaeten/biowissenschaften/ipmb/biologie/woelfl/Research.html. By introducing the program, we hope to encourage more systematic use of time-resolved assays and lead researchers to fully exploit their data.

No MeSH data available.


Related in: MedlinePlus

Time-resolved IC50 of MCF-7 cells exposed to Cisplatin.The oxygen level in the media of MCF-7 cells treated with different Cisplatin concentrations between days 2.2 and 5 (as depicted in Fig 3A) are fitted for each time point, as exemplified for day 2.50 (green), 2.89 (turquoise), 3.28 (blue), 3.66 (pink), 4.05 (red), 4.43 (yellow) (A). MicroM stands for micromolar. The IC50 over time (B) is then determined from the values of all the fits between days 2.5 and 4.7. The residual error of the fit is displayed as a grey shadow around each curve.
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pone.0131233.g004: Time-resolved IC50 of MCF-7 cells exposed to Cisplatin.The oxygen level in the media of MCF-7 cells treated with different Cisplatin concentrations between days 2.2 and 5 (as depicted in Fig 3A) are fitted for each time point, as exemplified for day 2.50 (green), 2.89 (turquoise), 3.28 (blue), 3.66 (pink), 4.05 (red), 4.43 (yellow) (A). MicroM stands for micromolar. The IC50 over time (B) is then determined from the values of all the fits between days 2.5 and 4.7. The residual error of the fit is displayed as a grey shadow around each curve.

Mentions: The four-parameter log-logistic model (Eq 2) is fitted to the averaged data for every time point between day 2.5 and 4.7 (Fig 4A). The parameter e, which is the IC50 value, is then plotted over time to display the time-resolved IC50 of Cisplatin (Fig 4B) which ranges from 80 μM at day 2.5, to 30μM at day 4.7. These values are in accordance with previously published data [15, 16] taking into account diverging protocol details (seeding density, Cisplatin solvant, viability test, time points). The longer the exposure to Cisplatin, the lower the concentration needed to reach 50% control viability. This decrease begins almost immediately, it is fast for short exposure times but levels out progressively, almost reaching a steady-state. Cisplatin has a rapid effect on cell respiration combined with a high cell permeability, its lack of stability over time could explain the levelling out.


Time-resolved cell culture assay analyser (TReCCA Analyser) for the analysis of on-line data: data integration--sensor correction--time-resolved IC50 determination.

Lochead J, Schessner J, Werner T, Wölfl S - PLoS ONE (2015)

Time-resolved IC50 of MCF-7 cells exposed to Cisplatin.The oxygen level in the media of MCF-7 cells treated with different Cisplatin concentrations between days 2.2 and 5 (as depicted in Fig 3A) are fitted for each time point, as exemplified for day 2.50 (green), 2.89 (turquoise), 3.28 (blue), 3.66 (pink), 4.05 (red), 4.43 (yellow) (A). MicroM stands for micromolar. The IC50 over time (B) is then determined from the values of all the fits between days 2.5 and 4.7. The residual error of the fit is displayed as a grey shadow around each curve.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4482264&req=5

pone.0131233.g004: Time-resolved IC50 of MCF-7 cells exposed to Cisplatin.The oxygen level in the media of MCF-7 cells treated with different Cisplatin concentrations between days 2.2 and 5 (as depicted in Fig 3A) are fitted for each time point, as exemplified for day 2.50 (green), 2.89 (turquoise), 3.28 (blue), 3.66 (pink), 4.05 (red), 4.43 (yellow) (A). MicroM stands for micromolar. The IC50 over time (B) is then determined from the values of all the fits between days 2.5 and 4.7. The residual error of the fit is displayed as a grey shadow around each curve.
Mentions: The four-parameter log-logistic model (Eq 2) is fitted to the averaged data for every time point between day 2.5 and 4.7 (Fig 4A). The parameter e, which is the IC50 value, is then plotted over time to display the time-resolved IC50 of Cisplatin (Fig 4B) which ranges from 80 μM at day 2.5, to 30μM at day 4.7. These values are in accordance with previously published data [15, 16] taking into account diverging protocol details (seeding density, Cisplatin solvant, viability test, time points). The longer the exposure to Cisplatin, the lower the concentration needed to reach 50% control viability. This decrease begins almost immediately, it is fast for short exposure times but levels out progressively, almost reaching a steady-state. Cisplatin has a rapid effect on cell respiration combined with a high cell permeability, its lack of stability over time could explain the levelling out.

Bottom Line: The functions of the program include data normalising and averaging, as well as smoothing and slope calculation, pin-pointing exact change time points.To illustrate the capabilities of the TReCCA Analyser, we performed on-line monitoring of dissolved oxygen in the culture media of the breast cancer cell line MCF-7 treated with different concentrations of the anti-cancer drug Cisplatin.The TReCCA Analyser is freely available at www.uni-heidelberg.de/fakultaeten/biowissenschaften/ipmb/biologie/woelfl/Research.html.

View Article: PubMed Central - PubMed

Affiliation: Institute for Pharmacy and Molecular Biotechnology, Heidelberg University, Heidelberg, Germany; Institute for Analytical Chemistry, Mannheim University of Applied Sciences, Mannheim, Germany.

ABSTRACT
Time-resolved cell culture assays circumvent the need to set arbitrary end-points and reveal the dynamics of quality controlled experiments. However, they lead to the generation of large data sets, which can represent a complexity barrier to their use. We therefore developed the Time-Resolved Cell Culture Assay (TReCCA) Analyser program to perform standard cell assay analyses efficiently and make sophisticated in-depth analyses easily available. The functions of the program include data normalising and averaging, as well as smoothing and slope calculation, pin-pointing exact change time points. A time-resolved IC50/EC50 calculation provides a better understanding of drug toxicity over time and a more accurate drug to drug comparison. Finally the logarithmic sensor recalibration function, for sensors with an exponential calibration curve, homogenises the sensor output and enables the detection of low-scale changes. To illustrate the capabilities of the TReCCA Analyser, we performed on-line monitoring of dissolved oxygen in the culture media of the breast cancer cell line MCF-7 treated with different concentrations of the anti-cancer drug Cisplatin. The TReCCA Analyser is freely available at www.uni-heidelberg.de/fakultaeten/biowissenschaften/ipmb/biologie/woelfl/Research.html. By introducing the program, we hope to encourage more systematic use of time-resolved assays and lead researchers to fully exploit their data.

No MeSH data available.


Related in: MedlinePlus