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xCELLigence RTCA ePacer

An innovative solution for maturing hiPSC Cardiomyocytes into a more predictive model.

The xCELLigence RTCA ePacer provides an easy and effective way to produce functionally mature hiPSC cardiomyocytes. Under precise and consistent electrical pacing conditions, the ePacer improves the maturation status of the hiPSC cardiomyocytes in just 2-3 weeks, with much less stress compared to existing pacing methods.

 

Figure: After being subjected to directed progressive electrical pacing using the xCELLigence RTCA ePacer, the hiPSC cardiomyocytes were able to demonstrate a positive force-frequency relationship and appropriate contractile responses to inotropes, significantly improved organized sarcomere structure, and proper gene expression.

Download the xCELLigence RTCA ePacer Brochure


Software

Tunable Pacing Function

Stimulus settings, such as pulse type, pulse intensity, pulse length, and stimulation duration can be easily selected and defined by the user.

Independent Pacing Settings for Individual Columns and Plates

Stimulus settings can be applied based on individual columns or across different plates.

Quick and Easy Way to Evaluate Cell Maturation Status Using Force-frequency Relationship Calculation

The built-in algorithm automatically generates force-frequency relationship curves before and after long-term pacing process.


Instrument

DescriptionCat. #
xCELLigence RTCA ePacer Bundle
xCELLigence RTCA ePacer Analyzer380601520
ePacer Station with 6 Cardio Cradles380601530
ePacer Station with 6 CardioECR Cradles380601540
ePacer Station with 3 Cardio Cradles and 3 CardioECR Cradles380601550
ePacer Station-Customer Configuration380601560

Consumables

DescriptionCat. #
E-Plate Cardio 96 (6 plates)300601050
E-Plate Cardio 96 (36 plates)300601060
E-Plate Cardio View 96 (6 plates)300601080
E-Plate Cardio View 96 (36 plates)300601090
E-Plate CardioECR 48 (6 Plates)300601110
E-Plate CardioECR 48 (36 Plates)300601120

Spec Sheets

Publications

Videos

Webinar: Electrical Field Stimulation for the Functional Maturation of hiPSC-Cardiomyocytes

Recorded on June 27, 2019

One of the biggest gaps to full utilization of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) is their inherent maturation status. In this webinar, Dr. Xiaoyu Zhang and Dr. T.K. Feaster discuss exciting new data from a novel high-throughput instrument that uses electrical conditioning to achieve functionally mature cardiomyocytes that can be used to assess excitation-contraction coupling and other orthogonal assays.

Watch Now

Attend this webinar to learn about:

  • Principle and protocol for electrical pacing of cardiomyocytes
  • Measurable characteristics of matured cardiomyocytes
  • A new instrument that allows high-throughput maturation
  • Implications for the future of cardiac research (workflow models)

Who should attend:

  • Scientists using human-induced pluripotent stem cell cardiomyocytes
  • Safety/tox assessment, drug discovery, cardiac disease modeling

Presenters

Presenter
Dr. Xiaoyu Zhang

Principal Scientist
ACEA Biosciences – a part of Agilent

Xiaoyu Zhang received her Ph.D. in cell molecular and developmental biology from the University of California, Riverside, USA, and conducted postdoctoral training and research at the Burnham Institute for Medical Research. Dr. Zhang joined ACEA Biosciences in 2011 where she has been leading the FDA CiPA studies, cardiac safety screening program and RTCA Cardio/CardioECR applications development.
Presenter
Dr. T.K. Feaster

Product Manager
Fujifilm Cellular Dynamics

Dr. Feaster received his Ph.D. in Pharmacology in 2015 from Vanderbilt University Medical Center for his work using human-induced pluripotent stem cell-derived cardiomyocytes to investigate cardiac excitation-contraction coupling. Dr. Feaster joined FCDI as a technical application scientist working in the applications group, where he served as the primary support liaison for cardiac products, as well as sharing responsibility for in-lab application development. In 2016, Dr. Feaster moved into a field application scientist role for FCDI’s toxicology and safety pharmacology business unit. Dr. Feaster is currently the product manager for the cardiac portfolio at FCDI.
xCELLigence RTCA ePacer Product Video

Videos: Product Overviews

xCELLigence RTCA ePacer Product Video - 

Videos: Research Presentations

Webinar: Electrical Field Stimulation for the Functional Maturation of hiPSC-Cardiomyocytes - Recorded on June 27, 2019 One of the biggest gaps to full utilization of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) is their inherent maturation status. In this webinar, Dr. Xiaoyu Zhang and Dr. T.K. Feaster discuss exciting new data from a novel high-throughput instrument that uses electrical conditioning to achieve functionally mature cardiomyocytes that … Continued

Recorded on June 27, 2019

One of the biggest gaps to full utilization of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) is their inherent maturation status. In this webinar, Dr. Xiaoyu Zhang and Dr. T.K. Feaster discuss exciting new data from a novel high-throughput instrument that uses electrical conditioning to achieve functionally mature cardiomyocytes that can be used to assess excitation-contraction coupling and other orthogonal assays.

Watch Now

Attend this webinar to learn about:

  • Principle and protocol for electrical pacing of cardiomyocytes
  • Measurable characteristics of matured cardiomyocytes
  • A new instrument that allows high-throughput maturation
  • Implications for the future of cardiac research (workflow models)

Who should attend:

  • Scientists using human-induced pluripotent stem cell cardiomyocytes
  • Safety/tox assessment, drug discovery, cardiac disease modeling

Presenters

Presenter
Dr. Xiaoyu Zhang

Principal Scientist
ACEA Biosciences – a part of Agilent

Xiaoyu Zhang received her Ph.D. in cell molecular and developmental biology from the University of California, Riverside, USA, and conducted postdoctoral training and research at the Burnham Institute for Medical Research. Dr. Zhang joined ACEA Biosciences in 2011 where she has been leading the FDA CiPA studies, cardiac safety screening program and RTCA Cardio/CardioECR applications development.
Presenter
Dr. T.K. Feaster

Product Manager
Fujifilm Cellular Dynamics

Dr. Feaster received his Ph.D. in Pharmacology in 2015 from Vanderbilt University Medical Center for his work using human-induced pluripotent stem cell-derived cardiomyocytes to investigate cardiac excitation-contraction coupling. Dr. Feaster joined FCDI as a technical application scientist working in the applications group, where he served as the primary support liaison for cardiac products, as well as sharing responsibility for in-lab application development. In 2016, Dr. Feaster moved into a field application scientist role for FCDI’s toxicology and safety pharmacology business unit. Dr. Feaster is currently the product manager for the cardiac portfolio at FCDI.

Videos: Tutorials

Videos: J.O.V.E. Video Articles

Videos: Testimonials

Applications Overview

The xCELLigence RTCA ePacer is adaptable and can easily integrate into your existing assay workflow. The view area on the E-Plate Cardio View 96 allows for compatibility with other optical assays.

With a focus of developing more functionally mature cardiomyocytes, the applications that the RTCA ePacer can be used for is vast.

Cardiac Drug Discovery

Innovate Your Approach to Drug Discovery

Dynamic and intricate monitoring of the excitation-contraction process of cardiomyocytes

The 5-year mortality rate for the over 23 million heart failure (HF) patients remains high, despite extensive efforts in development and investigation of therapeutic agents for the management HF. The physiological benefits of the treatments have often been coupled with unwanted side effects. Human induced pluripotent stem cell cardiomyocytes (hiPSC-CM) offer an exciting new model system to test both the efficacy and toxicity of new therapeutic approaches, including inotropic compounds which serve to modulate the force of cardiomyocyte contractility.

Assessment of Inotropic Compounds

Inotropic assays and other ex vivo whole organ assays are labor intensive, low throughput, and expensive. Alternatively, animal studies can also be conducted, but the actual predictivity of these assays and their translational potential to humans needs to be taken into account. Human iPSC-CM can provide a relevant model system for assessment of inotropic compounds. However, because of the relative immaturity of hiPSC-CM, assessment of inotropic compounds remain a challenge. ACEA Biosciences has developed an electrical pacing protocol which functionally matures hiPSC-CM (see maturity) enabling assessment of positive and negative inotropic compounds. Below is a summary table of inotropic compounds tested with electrically paced and functionally matured hiPSC-CM.

Figure 1: Summary of contractile responses of long-term electrically paced iCell CM2s to a panel of inotropic compounds, including 6 positive inotropes (+) and 1 negative inotrope (-)

Overcome hurdles in your research to find your next insight of compounds effecting the contractile process

Test Alt

Webinar: Evaluating Drug Induced Proarrhythmic Risk Using the CardioECR System

Dr. Haoyu Zeng describes his experience using the CardioECR system to screen drugs as part of the FDA CiPA initiative.

Watch Webinar
Jason Maynes Video

Webinar: Exploring the Role of iPSC-Cardiomyocytes in Drug Discovery and Safety Assessment

Dr. Kristina Green (Myokardia) and Dr. Armando Lagrutta (Merck) will describe new approaches aimed at enhancing assessment of hiPSC-CM functionality.

Watch Webinar
Test Alt

Publication: High throughput physiological screening of iPSC cardiomyocytes for drug development

A review of emerging technologies for high throughput measurements of cardiomyocyte physiology, and comments on using iPSC cardiomyocytes to model disease & drug discovery. 

Read More

Easy and Flexible Work Flow: Simply plate the cells, start acquiring data, and perform combination treatments & chronic dosing

Simplify your analysis – powerful software to help you elucidate information from data
Unique software monitors integrated ion channel activity, contractility, and longer term viability to elucidate adverse compound action.

Click to Learn More

 

See What Real-Time Cell Analysis Does For You

The xCELLigence CardioECR® system combines real-time impedance monitoring with both multielectrode array (MEA) technology and a pacing function for the evaluation of cardiomyocyte function along with integrated ion channel activity.

Click to Learn More

The xCELLigence® RTCA Cardio system uses non-invasive, label-free impedance monitoring to quantitatively evaluate cardiomyocyte health/function in real-time.

Click to Learn More

 

Are your hiPSC-CMs displaying neonatal gene expression profile? The xCELLigence RTCA CardioECR system has a powerful electrical pacing capability which can be used to functionally mature hiPSC-CM which display more adult-like functional profile and a more organized calcium handling machinery.

Click here to learn more


Training and Webinars

ACEA Biosciences offers webinars and free online training to help you find your next breakthrough

View WebinarsView App Notes

 

 


ACEA Biosciences – The Cell Analysis Company

At ACEA Biosciences, our goal is to apply innovative technologies to the analysis of cellular mechanism and response, making studies possible that were not even imaginable just a few years ago. As a global company that places high value on collaborative interactions, rapid delivery of solutions, and providing the highest level of quality, we strive to meet our customers’ needs by delivering innovative, flexible, and scalable solutions to advance the next breakthrough in elucidating mechanisms of the human body. ACEA real-time cell analysis and flow cytometry technologies are fueling groundbreaking advancements in life science research and molecular diagnostics.

Cardiomyocyte Functional Assays

Physiologically Relevant Cell Types with Real-Time Cell Analysis for More Predictive Assays

 

Continuous real-time cell analysis improves on the laborious end-point assays by providing complete uninterrupted cellular response over the course of your study. Monitor the functional aspects of human induced pluripotent stem cell cardiomyocyte (hiPSC-CM) or primary cardiomyocytes over the entire duration of the experiment for more comprehensive functional evaluation in the context of safety/toxicity assessment, screening of cardiac drugs, or evaluating molecular mechanisms of disease.

 


Cardio Applications


 Cardiac Safety ToxicityCardiac Drug DiscoveryCardiac Disease ModelingCardiomyocyte MaturationCiPA

Jason Maynes Video

Video: Studying pediatric cardiac diseases in vitro using xCELLigence CardioECR system

Dr. Jason Maynes, analyzing contractility and ion channel activity of diseased pediatric iPSC-CMs using the CardioECR system.

Watch Video
Test Alt

Webinar: Exploring the Role of iPSC-CM in Drug Discovery and Safety Assessment

Scientists from MyoKardia and Merck describe new approaches aimed at enhancing assessment of hiPSC-CM functionality

Watch Webinar
Test Alt

Brochure: The Cardio & CardioECR Systems

Read and learn more about the comprehensive solution for in vitro cardiac safety assessment and cardiovascular drug discovery.

Watch Webinar

 

Simplify your workflow. No cell labeling required. Simply seed your cells, monitor, and evaluate the mechanism, efficacy or toxicity response of pharmaceutical drugs in cardiomyocytes

 

See What Real-Time Cell Analysis Does For You

The xCELLigence CardioECR® system combines real-time impedance monitoring with both multielectrode array (MEA) technology and a pacing function for the evaluation of cardiomyocyte function along with integrated ion channel activity.

Click to Learn More

The xCELLigence® RTCA Cardio system uses non-invasive, label-free impedance monitoring to quantitatively evaluate cardiomyocyte health/function in real-time.

Click to Learn More

 

Training and Webinars

ACEA Biosciences offers webinars and free online training to help you find your next breakthrough

View WebinarsView App Notes

 

ACEA Biosciences – The Cell Analysis Company

At ACEA Biosciences, our goal is to apply innovative technologies to the analysis of cellular mechanism and response, making studies possible that were not even imaginable just a few years ago. As a global company that places high value on collaborative interactions, rapid delivery of solutions, and providing the highest level of quality, we strive to meet our customers’ needs by delivering innovative, flexible, and scalable solutions to advance the next breakthrough in elucidating mechanisms of the human body. ACEA real-time cell analysis and flow cytometry technologies are fueling groundbreaking advancements in life science research and molecular diagnostics.

Cardiomyocyte Maturation

Functional Maturation of hiPSC-CMs

Utility of hiPSC-CMs for Truly Predictive Assays

Human iPSC-CM’s inherent morphology, structural hallmarks, calcium handling mechanism, electrophysiology, and gene expression profile resemble neonatal cardiomyocytes which limits its application for safety/toxicity assessment, drug discovery and even disease modeling.

Long-term electrical pacing is quickly becoming the preferred method of choice to improve the maturation status of hiPSC-CMs in a consistent and scalable manner. Recent discoveries using an Integrated Pacing Function to pace hiPSC-CM indicate that the functionally mature hiPSC-CM provide a significantly improved model that can be used for various applications including safety/tox assessment, drug discovery, and cardiac disease modeling. The xCELLigence CardioECR system utilizes planar impedance electrodes to electrically pace hiPSC-CM and achieve reliable pacing of hiPSC-CM.

Long-term Electrical Pacing Improves Maturation of hiPSC-CM

Human iPSC-CM were electrically paced with the xCELLigence CardioECR system using escalating frequency of pacing over 3 weeks and then assessed for gene expression, protein expression, and functional assays.

Improve Gene Expression by Electrical Pacing
The expression of key maturation gene markers in non-paced (CTRL) and paced cardiomyocytes was determined by RT-PCR. The human fetal heart (FH) and adult left ventricle (ALV) were included as benchmarks of known level of maturation. The amount of RNA was normalized to the expression of GAPDH. Gene expression level was presented as a fold change to the human fetal heart. The data are presented as mean± SD, N=4. (* P< 0.05).
Improve Protein Expression by Electrical Pacing
 Protein expression level of sarcomeric α-actinin, myosin heavy chain, cTNT, SERCA2a and PLN in paced and non-paced cells were determined by western blot analysis, which was quantified using protein to GAPDH ratio and further normalized to the non-paced cardiomyocytes. The data are presented as mean ± SD, N=4. (* P< 0.05).
Improve Contractile Response by Electrical Pacing
 Isoproterenol treatment in CTRL and paced cardiomyocytes (a-d). Non-paced and paced cells were treated with ISO at 10, 100 and 1000 nM. The averaged percentage change of beating amplitude (a) and beating rate (b) to the time-matched DMSO control were calculated in non-paced/CTRL (in blue) and paced cells (in orange) at 30 min after ISO exposure. The data are presented as mean ± SD, N=3. The IMP waveforms were continuously measured for 30 s before and after drug addition, which were further calculated to averaged single waveforms (C). Representative averaged IMP waveforms obtained before (in black) and after ISO addition (paced cells, orange trace). (See Drug Discovery)

Human iPSC-CMs possess an inherent negative impedance amplitude-frequency relationship which is reversed after electrical pacing. 

Overcome cardiomyocyte neonatal phenotype expression to obtain truly predictive assays
Safety/Toxicity
Drug Discovery
Disease Models

Training and Webinars

ACEA Biosciences offers webinars and free online training to help you find your next breakthrough

View WebinarsView App Notes

 

 


ACEA Biosciences – The Cell Analysis Company

At ACEA Biosciences, our goal is to apply innovative technologies to the analysis of cellular mechanism and response, making studies possible that were not even imaginable just a few years ago. As a global company that places high value on collaborative interactions, rapid delivery of solutions, and providing the highest level of quality, we strive to meet our customers’ needs by delivering innovative, flexible, and scalable solutions to advance the next breakthrough in elucidating mechanisms of the human body. ACEA real-time cell analysis and flow cytometry technologies are fueling groundbreaking advancements in life science research and molecular diagnostics.

Cardiotoxicity & Drug Screening

Is Your Current Technology Predictive?

xCELLigence® Cardio and CardioECR instruments provide the answers:

  • Superb Predictivity: Easily screen and quickly identify short-term and long-term cardiac toxicity early in drug development. (Figure 1)
  • Easy and Flexible Work Flow: Simply plate the cells, start acquiring data, and perform combination treatments and chronic dosing.
  • Powerful Multiplexing: Simultaneous readout of cardiomyocyte contractility, integrated ion channel activity (Figure 2), and viability.
  • Full Control of Beating Rate Enables Functional Maturation: Directed pacing feature improves functionality of iPSC cardiomyocytes (Figure 3) and response to inotropic compounds.
Request a Demo

 

Pharmacological assessment of ion channel inhibitors using iCell cardiomyocytesFigure 1. Pharmacological assessment of ion channel inhibitors using iCell cardiomyocytes. iCell cardiomyocytes exhibiting consistent and robust beating activity (14 days post-seeding) were treated with multiple concentrations of each compound for up to 24 hours. Compound-induced arrhythmia was recorded by the xCELLigence RTCA Cardio instrument. 

 

Cardiac excitation-contraction couplingFigure 2. Cardiac excitation-contraction coupling. The xCELLigence CardioECR instrument simultaneously measures electrophysiological signals and contractility. Blebbistatin, a myosin inhibitor, does not inhibit the ion-channel signal of treated cells (green), but greatly impairs the mechanical contraction and beating (red). Being able to monitor this excitation-contraction coupling relationship provides a complete picture for safety assessment of compounds during drug development.

 

Beating amplitude and Beating Rate Relations over time

Figure 3. Beating amplitude and Beating Rate Relations over time. iCell cardiomyocytes exhibiting consistent and robust beating and field potential signals were either submitted to directed progressive electrical stimulation (Paced) using the xCELLigence CardioECR instrument or cultured without electrical stimulation (Spontaneous) for 3 weeks. The Beating amplitude and Beating Rate relationship was determined weekly.

Download the Functional Maturation of iPSC Cardiomyocytes Flyer


Cardiotoxicity & Drug Screening Supporting Information:

  • Compatible xCELLigence® Systems
xCELLigence RTCA CardioxCELLigence RTCA CardioECR
RTCA CardioRTCA CardioECR
Throughput1×96 wells1×48 wells
ReadoutImpedance onlyImpedance and Field Potential
Sampling RateImpedance: 12.9 msImpedance: 1 ms
Field Potential: 10 kHz
  • Cardiotoxicity Publications:
  1. Estimating the Risk of Drug-induced Proarrhythmia Using Human Induced Pluripotent Stem Cell-derived Cardiomyocytes. Guo L, Abrams RM, Babiarz JE, Cohen JD, Kameoka S, Sanders MJ, Chiao E, Kolaja KL. Toxicol Sci. 2011 Sep;123(1):281-9. (Hoffmann-La Roche, Nutley, USA)
  2. Impedance-based Detection of Beating Rhythm and Proarrhythmic Effects of Compounds on Stem Cell-derived Cardiomyocytes. Jonsson MK, Wang QD, Becker B. Assay Drug Dev Technol. 2011 Dec;9(6):589-99. (Roche R&D, Sweden)
  3. Functional Cardiotoxicity Profiling and Screening Using the xCELLigence RTCA Cardio System. Xi B, Wang T, Li N, Ouyang W, Zhang W, Wu J, Xu X, Wang X, Abassi YA. J Lab Autom. 2011 Dec;16(6):415-21. (ACEA Biosciences, Inc., USA)
  4. In vitro Model for Assessing Arrhythmogenic Properties of Drugs Based on high-resolution Impedance Measurements. Nguemo F, Šarić T, Pfannkuche K, Watzele M, Reppel M, Hescheler J. Cell Physiol Biochem. 2012;29(5-6):819-32. (University of Cologne, Germany)
  5. Dynamic Monitoring of Beating Periodicity of Stem Cell-derived Cardiomyocytes as a Predictive Tool for Preclinical Safety Assessment. Abassi YA, Xi B, Li N, Ouyang W, Seiler A, Watzele M, Kettenhofen R, Bohlen H, Ehlich A, Kolossov E, Wang X, Xu X. Br J Pharmacol. 2012 Mar;165(5):1424-41. (ACEA Biosciences, Inc.,USA)
  6. Drug-induced Functional Cardiotoxicity Screening in Stem Cell-derived Human and Mouse Cardiomyocytes: Effects of Reference Compounds. Himmel HM. J Pharmacol Toxicol Methods. 2013 Jul-Aug;68(1):97-111. (Bayer Pharma AG, Germany)
  7. Refining the Human iPSC Cardiomyocyte Arrhythmic Risk Assessment Model. Guo L, Coyle L, Abrams RM, Kemper R, Chiao ET, Kolaja KL. Toxicol Sci. 2013 Dec;136(2):581-94. (Hoffmann-La Roche, Inc., Nutley, USA)
  8. The Proliferative and Chronotropic Effects of Brillantaisia Nitens Lindau (Acanthaceae) Extracts on Pluripotent Stem Cells and Their Cardiomyocytes Derivatives. Nembo EN, Dimo T, Bopda OS, Hescheler J, Nguemo F. J Ethnopharmacol. 2014 Oct 28;156:73-81. (University of Cologne, Germany)
  9. Chapter 16: Label-Free Impedance Measurements for Profiling Drug- Induced Cardiotoxicity. Nguemo F, Semmler J, Hescheler J. Label-Free Biosensor Methods in Drug Discovery 2015 (University of Cologne, Germany)

 

Preclinical In Vitro Cardiac Toxicity

Contractile Activity + Electrophysiological Activity + Viability -> Real Time Cell Analysis of Toxicity

Cardiomyocytes for the predictive assessments of potential cardiotoxic side effects

Significant numbers of drug development projects are terminated in the late preclinical and early clinical stages due to cardiac liability issues. The last two decades has witnessed increased withdrawals or issued safety warnings due to cardiotoxicity. Approximately 1/3 of all drugs withdrawn during 1990-2006 were associated with Torsades de Pointes (TdP). One of the main challenges in preclinical cardiac safety assessment has been the lack of a predictive and biologically relevant model system available in sufficiently high quantities to be used for screening of cardiotoxic and pro-arrhythmic drugs, especially during the hit to lead or lead optimization stage. Recent advances in differentiating embryonic and induced-pluripotent stems cells have created physiologically relevant and disease relevant model systems for preclinical safety assessment of compounds.

Assessment of Contractile Related Toxicities

The existing methods for assessment of contractile activities, such as Langendorff heart assays and other ex vivo whole organ assays are labor intensive, low throughput, and expensive. Alternatively, animal studies can also be conducted, but the predictivity of these assays and translational potential to humans is always an important consideration in addition to the ethical considerations of using animals. Functional monitoring of cardiomyocyte contractility using impedance readout (see figure below) provides incisive information about compound effects on the contractile machinery.

Figure 1: Impedance-based contraction signal of different types of hiPSC-CM.

 

Case study: Blebbistatin, a myosin heavy chain inhibitor, decouples excitation and contraction coupling.

  • Blebbistatin reduced beating amplitude in a dose-dependent manner, but did not alter the field potential signal even after exposure to 10 µM blebbistatin.

(A) 5 seconds of beating amplitude traces obtained from iCell cardiomyocytes before (black traces)  and after (red traces) blebbistatin treatment at 0.01, 1, and 10 µM. The compound addition was performed in one dose one well fashion. (B) 5 seconds of FP traces obtained before (black trace) and after 10 µM of blebbistatin treatment.

Assessment of Ion Channel Toxicities

Assays that more directly assess possible pro-arrhythmic effects of compounds utilize whole animal hearts or Purkinje fibers, and are designed to assess action potential duration. While these assays are considered to be more predictive of arrhythmia, they have higher negative predictivity rate and can be low throughput and technically demanding. Human iPSC-CMs provide a physiologically relevant model that can be used to assess potential toxicity of pharmaceutical compounds or integrated ion channel activity.

Figure 2: Field potential signal of hiPSC-CM.

Assessment of Acute Toxicities:

1. Na+ channel>Mexiletine, a Na+ channel blocker, induced reduction of field potential (FP) spike, suggesting the blockade of sodium channel.

5 seconds of impedance (IMP) (A) and field potential (FP) (B) signals of iCell CMs before (black traces) and after (red traces) exposure to 10 µM Mexiletine. The raw IMP and FP traces were then further calculated to single averaged waveforms (C and D).

2. Ca2+ channelsIsradipine, a Ca2+channel blocker, induced field potential duration (FPD) shortening and decrease in IMP amplitude/beating amplitude (BAmp).

5 seconds of impedance (IMP) (A) and field potential (FP) (B) signals of iCell cardiomyocytes before (black traces) and after (red traces) exposure to 0.01 µM isradipine. The raw IMP and FP traces were then further calculated to single averaged waveforms (C and D).

3. hERG channelE4031, a hERG channel blocker, induced EAD-like FP signal and arrhythmic beating on beating amplitude traces.

5 seconds of impedance (beating amplitude) (A) and field potential (FP) (B) signals of iCell cardiomyocytes before (black traces) and after (red traces) exposure to 0.03 µM E4031. The overlay of IMP and FP traces (C) 30 min post-drug.

4. Multi-ion channelFlecainide: a multichannel blocker, which inhibits Na+ and hERG channels. Flecainide induced FP spike reduction and EAD-like (notch and ectopic beat) on FP traces and arrhythmic beating on IMP traces.

5 seconds of impedance (IMP) (A) and field potential (FP) (B) signals of iCell cardiomyocytes before (black traces) and after (red traces) exposure to 3µM flecainide. (C)The overlay of IMP and FP traces (C) 30 min post-drug. (D) Magnified in FP trace.

Long-term Ion Channel Toxicities:

Pentamidine, a hERG channel trafficking inhibitor, showed delayed FPD and contractility period duration.

(A) 15 seconds of IMP traces obtained from iCell cardiomyocytes before and after pentamidine treatment at the indicated time points. (B) The expanded FP traces recorded before and after 10 µM pentamidine treatment at the indicated time points. (C) Time course of FPD measurement over the course of 10 µM pentamidine treatment.

Assessment of Structural Related Toxicities

Chemotherapeutic drugs, such as anthracyclines and certain kinase inhibitors, are known to compromise the structural integrity of cardiomyocytes. Identifying and delineating these types of toxicities are an important component of overall cardiotoxicity assessment of pharmaceutical compounds. While there may be laborious orthogonal end-point assays that can provide information regarding the impact of pharmaceutical compounds on structural integrity of cardiac myocytes, some effects may be transient and missed by end-point analysis. The xCELLigence CardioECR system is the only assay or platform that provide both structural integrity and viability information, while simultaneously providing data about ion channel activity and contractility. The totality of the three types of toxicities makes the CardioECR a unique platform well positioned for early cardiotoxicity screening.


Assessment of Anthracycline Toxicity:

Doxorubicin, an anthracycline and chemotherapy drug, is known to cause cardiac toxicity

(A) Overall cell index change over the course of 72 hour treatment. (B) 15 seconds of impedance traces obtained from iCell cardiomyocytes before and after doxorubicin treatment at the indicated time points.

  • Doxorubicin induced overall cell index decrease in a dose and time-dependent manner, suggesting a loss of cell viability and/or structural damage to the cells.
  • Doxorubicin also affected the contractility of cardiomyocytes in a dose and time dependent manner, which may be related to the loss of viability.

Assessment of Kinase Inhibitor Toxicity:

Sorafenib is a well-studied cardiotoxic compound that’s known to cause myocardial infarction and ischaemia.

  • Sorafenib induced overall cell index decrease in a dose and time-dependent manner. It caused cell quiescence at 20 µM

(A) Overall cell index change over the course of 48-hour treatment. (B) 15 seconds of impedance traces obtained from iCell cardiomyocytes before and after sorafenib treatment at the indicated time points. 

Overcome hurdles in your research to find your next insight of your therapeutic compound interaction

Ncardia webinar

Webinar: Rethinking Translational Predictivity using HiPSC-Derived Models

A Retrospective Analysis for Cardiac Liabilities, a webinar hosted by Ncardia and ACEA Biosciences.

Watch Webinar
Astrazeneca webinar

Webinar: Unraveling Kinase Inhibitor Cardiotoxicity with Cardiomyocyte Impedance Assays

Dr. Sarah Lamore (AstraZeneca) presents her research utilizing cellular impedance measurement of beating cardiomyocytes to deconvolute kinase inhibitor cardiotoxicity. 

Watch Webinar
cardiac safety paper

Publication: A Tool for Cardiac Safety Assessment

Multi-parametric assessment of cardiomyocyte excitation-contraction coupling using impedance and field potential recording.

Read More

Easy and Flexible Work Flow: Simply plate the cells, start acquiring data, and perform combination treatments & chronic dosing

Simplify your analysis – powerful software to help you elucidate information from data
Unique software monitors integrated ion channel activity, contractility, and longer term viability to elucidate adverse compound action.

Click to Learn More

 

Are you collecting all the needed data to make a true safety assessment?

The xCELLigence CardioECR® system combines real-time impedance monitoring with both multielectrode array (MEA) technology and a pacing function for the evaluation of cardiomyocyte function along with integrated ion channel activity.

Click to Learn More

The xCELLigence® RTCA Cardio system uses non-invasive, label-free impedance monitoring to quantitatively evaluate cardiomyocyte health/function in real-time.

Click to Learn More

Are your hiPSC-CMs displaying neonatal gene expression profile? Pacing hiPSC-CM under increasing pacing frequencies leading to functionally mature hiPSC-CM which display more adult-like gene expression profile.

Click here to learn more


Training and Webinars

ACEA Biosciences offers webinars and free online training to help you find your next breakthrough

View WebinarsView App Notes

 

 


ACEA Biosciences – The Cell Analysis Company

At ACEA Biosciences, our goal is to apply innovative technologies to the analysis of cellular mechanism and response, making studies possible that were not even imaginable just a few years ago. As a global company that places high value on collaborative interactions, rapid delivery of solutions, and providing the highest level of quality, we strive to meet our customers’ needs by delivering innovative, flexible, and scalable solutions to advance the next breakthrough in elucidating mechanisms of the human body. ACEA real-time cell analysis and flow cytometry technologies are fueling groundbreaking advancements in life science research and molecular diagnostics.

Predictive Toxicology: Cardiotoxicity

Drug toxicity is a pervasive issue in pharmaceutical drug development that needs to be mitigated. Failure to recognize such toxicities during drug development can pose not only great risks to the target population but also tremendous financial burdens and legal ramifications to drug developers in the event of drug withdrawals from the market.

One-third of all drugs withdrawn between 1990 and 2006 had been associated with cardiotoxicity, particularly ventricular arrhythmias, also known commonly as “Torsade de Pointes” (TdP). The xCELLigence® RTCA Cardio and CardioECR instruments monitor different functionalities of human (iPSC) and rodent cardiomyocyte including contractility, electrophysiology and viability in medium throughput manner. The CardioECR system is also capable of direct electrical pacing of cardiomyocytes, making it possible to assess compounds for their modulation of the force of contraction. This technology is currently being evaluated as part of the FDA Comprehensive Pro-Arrhythmia Assay (CiPA) initiative.


Cardiotoxicity Supporting Information:

  • Compatible xCELLigence® Systems
xCELLigence RTCA CardioxCELLigence RTCA CardioECR
RTCA CardioRTCA CardioECR
Throughput1×96 wells1×48 wells
ReadoutImpedance onlyImpedance and Field Potential
Sampling RateImpedance: 12.9 msImpedance: 1 ms
Field Potential: 10 kHz
  • Cardiotoxicity Publications:
  1. Estimating the Risk of Drug-induced Proarrhythmia Using Human Induced Pluripotent Stem Cell-derived Cardiomyocytes. Guo L, Abrams RM, Babiarz JE, Cohen JD, Kameoka S, Sanders MJ, Chiao E, Kolaja KL. Toxicol Sci. 2011 Sep;123(1):281-9. (Hoffmann-La Roche, Nutley, USA)
  2. Impedance-based Detection of Beating Rhythm and Proarrhythmic Effects of Compounds on Stem Cell-derived Cardiomyocytes. Jonsson MK, Wang QD, Becker B. Assay Drug Dev Technol. 2011 Dec;9(6):589-99. (Roche R&D, Sweden)
  3. Functional Cardiotoxicity Profiling and Screening Using the xCELLigence RTCA Cardio System. Xi B, Wang T, Li N, Ouyang W, Zhang W, Wu J, Xu X, Wang X, Abassi YA. J Lab Autom. 2011 Dec;16(6):415-21. (ACEA Biosciences, Inc., USA)
  4. In vitro Model for Assessing Arrhythmogenic Properties of Drugs Based on high-resolution Impedance Measurements. Nguemo F, Šarić T, Pfannkuche K, Watzele M, Reppel M, Hescheler J. Cell Physiol Biochem. 2012;29(5-6):819-32. (University of Cologne, Germany)
  5. Dynamic Monitoring of Beating Periodicity of Stem Cell-derived Cardiomyocytes as a Predictive Tool for Preclinical Safety Assessment. Abassi YA, Xi B, Li N, Ouyang W, Seiler A, Watzele M, Kettenhofen R, Bohlen H, Ehlich A, Kolossov E, Wang X, Xu X. Br J Pharmacol. 2012 Mar;165(5):1424-41. (ACEA Biosciences, Inc.,USA)
  6. Drug-induced Functional Cardiotoxicity Screening in Stem Cell-derived Human and Mouse Cardiomyocytes: Effects of Reference Compounds. Himmel HM. J Pharmacol Toxicol Methods. 2013 Jul-Aug;68(1):97-111. (Bayer Pharma AG, Germany)
  7. Refining the Human iPSC Cardiomyocyte Arrhythmic Risk Assessment Model. Guo L, Coyle L, Abrams RM, Kemper R, Chiao ET, Kolaja KL. Toxicol Sci. 2013 Dec;136(2):581-94. (Hoffmann-La Roche, Inc., Nutley, USA)
  8. The Proliferative and Chronotropic Effects of Brillantaisia Nitens Lindau (Acanthaceae) Extracts on Pluripotent Stem Cells and Their Cardiomyocytes Derivatives. Nembo EN, Dimo T, Bopda OS, Hescheler J, Nguemo F. J Ethnopharmacol. 2014 Oct 28;156:73-81. (University of Cologne, Germany)
  9. Chapter 16: Label-Free Impedance Measurements for Profiling Drug- Induced Cardiotoxicity. Nguemo F, Semmler J, Hescheler J. Label-Free Biosensor Methods in Drug Discovery 2015 (University of Cologne, Germany)