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2D Human Induced Pluripotent Stem Cell-derived Cardiomyocyte Cardiac Contractility Modulation Tool: 2D hiPSC-CM CCM Tool

Catalog of Regulatory Science Tools to Help Assess New Medical Devices 

 

This regulatory science tool presents a lab method to assist with evaluating and quantifying contractile response of non-excitatory electrical stimulation signals including CCM in vitro using 2D human cardiomyocyte models.

 

Technical Description

The primary objective of the 2D human induced pluripotent stem cell-derived cardiomyocyte (hiPSC-CM) Cardiac Contractility Modulation (CCM) Tool is to assist investigators with evaluating and quantifying contractile response of non-excitatory electrical stimulation signals including CCM in vitro using 2D human cardiomyocyte models. The 2D hiPSC-CM CCM Tool is a method and best practices package that includes detailed characterization, step-by-step protocols, and instructional video for healthy and/or diseased hiPSC-CM models in monoculture and co-culture.

The core elements of this tool include the ability to perform in vitro CCM contractility assays in 2D hiPSC models:

Step-by-Step Protocol and Instructional Video.

Intended Purpose

The 2D hiPSC-CM CCM Tool enables the user to rapidly generate the appropriate 2D in vitro environment to perform CCM contractility studies at the bench and to evaluate the standard CCM pulse parameters in various patient-specific hiPSC-CM models. This is important for safety and performance assessment and elucidation of molecular mechanisms of non-excitatory electrical stimulations. Moreover, this tool facilitates early-stage validation, de-risking product development and decision making through a more clinically relevant model. In addition, this tool enables physiological relevant bench testing using commercially available human cells bridging the gap between hiPSCs and medical device testing. This tool is a simple, easy, do-it-yourself package that uses standard laboratory equipment and technical capabilities relative to complex culture models including systems such as 3D human engineered cardiac tissues (ECTs).

This tool will enable less-experienced hiPSC-CM users to evaluate human contractile response in a reduced system. It is intended to be used by medical device developers, academia, and contract research organizations (CROs). This tool provides instantaneous feedback on whether the standard CCM parameters produce a contractile response in human cardiomyocytes from various healthy and disease backgrounds, as well as investigation of peripheral nervous system control over myocardial function in a nonclinical setting.

The 2D hiPSC-CM tool can be extended to various cardiac electrophysiological medical devices that deliver electrical stimulation to human myocardium through electrodes. Further, the 2D hiPSC-CM CCM tool can only evaluate the acute (i.e., seconds) effects of such stimulation.

Testing

Described in peer-reviewed publications:

  • Feaster, T. K., Casciola, M., Narkar, A., and Blinova, K. (2021). Acute effects of cardiac contractility modulation on human induced pluripotent stem cell-derived cardiomyocytes. Physiol. Rep. 9, e15085. doi:10.14814/phy2.15085
    • This publication describes the experimental details for maintenance and culture of 2D hiPSC-CMs monolayers including the experimental conditions necessary to elucidate nonexcitatory electrical stimulation (i.e., CCM) response under these conditions (i.e., flexible substrate and submaximal extracellular calcium). Moreover, the methodology to evaluate contractile properties is described as well as additional cardiac excitation-contraction coupling readouts including electrophysiology (i.e., action potential) and intracellular calcium handling. Demonstration of pharmacological characterization is presented with β-adrenergic antagonist and extracellular calcium challenge to evaluate potential mechanisms. Detailed description of the electrical set up, pulse parameters and numerical electric field modeling are provided.
  • Narkar, A., Feaster, T. K., Casciola, M., & Blinova, K. (2022). Human in vitro neurocardiac coculture (ivNCC) assay development for evaluating cardiac contractility modulation. Physiological Reports, 10, e15498. doi:10.14814/phy2.15498
    • This publication describes the experimental details for maintenance and culture of 2D hiPSC-CM (cardiomyocyte) and hiPSC-MN (Motor Neuron) monolayers (i.e., ivNCC, in vitro neurocardiac co-culture). Contractile response to nonexcitatory electrical stimulation (i.e., CCM) is described to elucidate cardiac-neuronal interplay and peripheral nervous system contribution. Detailed description of the electrical set up and pulse parameters are provided. Demonstration of pharmacological characterization is presented with, muscarinic agonist and β-adrenergic antagonist, to demonstrate functional coupling and elucidate potential mechanisms, respectively.
  • Feaster, T. K., Casciola, M., Narkar, A., Blinova, K. (2022). Evaluation of Cardiac Contractility Modulation Therapy in 2D Human Stem Cell-Derived Cardiomyocytes. J. Vis. Exp. (190), e64848, doi:10.3791/64848.
    • This publication describes the experimental details for maintenance and culture of 2D hiPSC-CMs monolayers as well as the application of nonexcitatory electrical stimulation (i.e., CCM). Step-by-step video and instructional protocol are provided. Likewise, the extension of CCM stimulation to a patient-specific disease model of Dilated Cardiomyopathy (DCM) is demonstrated.

Limitations

Device-specific verification, validation and optimization will be required to extend this tool to additional electrophysiological device signals. In future work the tool will be expanded to various electrophysiological non-excitatory electrical stimulation signals (i.e., pulse delay, pulse duration, pulse amplitude, and pulse number) in 3D microphysiological systems including 3D human engineered cardiac tissues (ECTs) to evaluate acute and chronic contractile response in vitro. This tool is not developed to replace in vivo animal models or clinical studies.

Supporting Documentation

Full technical description of the tool, test conditions, and experimental data are provided in:

Relevant Publications:

  • Feaster, T. K., Casciola, M., Narkar, A., and Blinova, K. (2021). Acute effects of cardiac contractility modulation on human induced pluripotent stem cell-derived cardiomyocytes. Physiol. Rep. 9, e15085. doi:10.14814/phy2.15085
  • Narkar, A., Feaster, T. K., Casciola, M., & Blinova, K. (2022). Human in vitro neurocardiac coculture (ivNCC) assay development for evaluating cardiac contractility modulation. Physiological Reports, 10, e15498. doi:10.14814/phy2.15498
  • Feaster, T. K., Casciola, M., Narkar, A., Blinova, K. (2022). Evaluation of Cardiac Contractility Modulation Therapy in 2D Human Stem Cell-Derived Cardiomyocytes. J. Vis. Exp. (190), e64848, doi:10.3791/64848

Related Work:

  • Blinova, K., Stohlman, J., Krauthamer, V., Knapton, A., Bloomquist, E., and Gray, R. A. (2014). Acute effects of nonexcitatory electrical stimulation during systole in isolated cardiac myocytes and perfused heart. Physiol. Rep. 2, e12106. doi:10.14814/phy2.12106
  • Blinova, K., Dang, Q., Millard, D., Smith, G., Pierson, J., Guo, L., et al. (2018). International multisite study of human-induced pluripotent stem cell-derived cardiomyocytes for drug proarrhythmic potential assessment. Cell. Rep. 24, 3582–3592. doi:10.1016/j.celrep.2018.08.079
  • Gintant, G., Kaushik, E. P., Feaster, T., Stoelzle-Feix, S., Kanda, Y., Osada, T., et al. (2020). Repolarization studies using human stem cell-derived cardiomyocytes: Validation studies and best practice recommendations. Regul. Toxicol. Pharmacol. 117, 104756. doi:10.1016/j.yrtph.2020.104756

Contact

For more information:

Appendix A:

For technical details regarding cell culture (monoculture), electrical apparatus, and stimulation:

Feaster, T. K., Casciola, M., Narkar, A., Blinova, K. (2022). Evaluation of Cardiac Contractility Modulation Therapy in 2D Human Stem Cell-Derived Cardiomyocytes. J. Vis. Exp. (190), e64848, doi:10.3791/64848

This publication describes the experimental details for maintenance and culture of 2D hiPSC-CMs monolayers as well as the application of nonexcitatory electrical stimulation (i.e., CCM). Step-by-step video and instructional protocol are provided. Likewise, the extension of CCM stimulation to a patient-specific disease model of Dilated Cardiomyopathy (DCM) is demonstrated.

For technical details regarding cell culture (co-culture), electrical apparatus, and stimulation:

Narkar, A., Feaster, T. K., Casciola, M., & Blinova, K. (2022). Human in vitro neurocardiac coculture (ivNCC) assay development for evaluating cardiac contractility modulation. Physiological Reports, 10, e15498. doi:10.14814/phy2.15498

This publication describes the experimental details for maintenance and culture of 2D hiPSC-CM (cardiomyocyte) and hiPSC-MN (Motor Neuron) monolayers (i.e., ivNCC, in vitro neurocardiac co-culture). Contractile response to nonexcitatory electrical stimulation (i.e., CCM) is described to elucidate cardiac-neuronal interplay and peripheral nervous system contribution. Detailed description of the electrical set up and pulse parameters are provided. Demonstration of pharmacological characterization is presented with, muscarinic agonist and β-adrenergic antagonist, to demonstrate functional coupling and elucidate potential mechanisms, respectively.

For technical details regarding cell culture (monoculture), electrical apparatus, and stimulation:

Feaster, T. K., Casciola, M., Narkar, A., and Blinova, K. (2021). Acute effects of cardiac contractility modulation on human induced pluripotent stem cell-derived cardiomyocytes. Physiol. Rep. 9, e15085. doi:10.14814/phy2.15085

This publication describes the experimental details for maintenance and culture of 2D hiPSC-CMs monolayers including the experimental conditions necessary to elucidate nonexcitatory electrical stimulation (i.e., CCM) response under these conditions (i.e., flexible substrate and submaximal extracellular calcium). Moreover, the methodology to evaluate contractile properties is described as well as additional cardiac excitation-contraction coupling readouts including electrophysiology (i.e., action potential) and intracellular calcium handling. Demonstration of pharmacological characterization is presented with β-adrenergic antagonist and extracellular calcium challenge to evaluate potential mechanisms. Detailed description of the electrical set up, pulse parameters and numerical electric field modeling are provided.