Functional and Pharmacological Analysis of Cardiomyocytes Differentiated from Human Peripheral Blood Mononuclear-Derived Pluripotent Stem Cells

Riedel, Michael; Jou, Chuanchau J.; Lai, Shuping; Lux, Robert L.; Moreno, Alonso P.; Spitzer, Kenneth W.; Christians, Elizabeth; Tristani‐Firouzi, Martin; Benjamin, Ivor J.

doi:10.1016/j.stemcr.2014.04.017

Cited by 39 publications

(38 citation statements)

References 47 publications

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“…The mean conduction velocity was 4.6 Ϯ 1.2 cm/s (n ϭ 5 independent experiments). These values are similar to the conduction velocities we recently reported in PBMC-derived iPSC-CMs measured using multielectrode array recordings (12). These proof-of-principle experiments highlight the ability to directly measure repolarization (i.e., APD 90 ) with the voltagesensitive dye rather than rely on estimates of repolarization based on the local field potential.…”

Section: Stability Of Di-4-anbdqbs Optical Ap Recordings and Photodynsupporting

confidence: 86%

“…PBMCs were isolated using standard techniques and reprogrammed by infecting cells with lentiviruses expressing reverse tetracycline-controlled transactivator, octamer-binding transcription factor 4, Kruppel-like factor 4, sex-determining region Y-box 2, and c-Myc genes from a polycistronic cassette (pHAGE2-TetONminiCMV-hSTEMCCA) (14,15), as recently described in Ref. 12. Infected blood cells were transferred onto mouse embryonic fibroblast (MEF) feeder cells 3 days postinfection and cultured for additional 7 days in MEF medium supplemented with 10 ng/ml human basic (b)FGF, 5 mg/ml ascorbic acid, and 2 mg/ml doxycycline.…”

Section: Methodsmentioning

confidence: 99%

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A near-infrared fluorescent voltage-sensitive dye allows for moderate-throughput electrophysiological analyses of human induced pluripotent stem cell-derived cardiomyocytes

López-Izquierdo

Warren

Riedel

et al. 2014

American Journal of Physiology-Heart and Circulatory Physiology

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-Human induced pluripotent stem cell-derived cardiomyocyte (iPSC-CM)-based assays are emerging as a promising tool for the in vitro preclinical screening of QT interval-prolonging side effects of drugs in development. A major impediment to the widespread use of human iPSC-CM assays is the low throughput of the currently available electrophysiological tools. To test the precision and applicability of the near-infrared fluorescent voltage-sensitive dye 1-(4-sulfanatobutyl)-4-{␤[2-(di-n-butylamino)-6-naphthyl]butadienyl}quinolinium betaine (di-4-ANBDQBS) for moderate-throughput electrophysiological analyses, we compared simultaneous transmembrane voltage and optical action potential (AP) recordings in human iPSC-CM loaded with di-4-ANBDQBS. Optical AP recordings tracked transmembrane voltage with high precision, generating nearly identical values for AP duration (AP durations at 10%, 50%, and 90% repolarization). Human iPSC-CMs tolerated repeated laser exposure, with stable optical AP parameters recorded over a 30-min study period. Optical AP recordings appropriately tracked changes in repolarization induced by pharmacological manipulation. Finally, di-4-ANBDQBS allowed for moderate-throughput analyses, increasing throughput Ͼ10-fold over the traditional patchclamp technique. We conclude that the voltage-sensitive dye di-4-ANBDQBS allows for high-precision optical AP measurements that markedly increase the throughput for electrophysiological characterization of human iPSC-CMs. optical action potential; preclinical drug screening; QT prolongation; acquired long QT syndrome; stem cell model RECENT ADVANCES in induced pluripotent stem cell (iPSC) technology now allow for the routine derivation of patient-and disease-specific human iPSC cardiomyocyte (iPSC-CM) models of cardiovascular disease. The impact of this technology has far-reaching implications, ranging from drug discovery, preclinical drug screening, mechanistic understanding of disease processes, and approaches for personalized medicine. Human iPSC-CM-based assays are emerging as a promising tool for the in vitro preclinical screening of QT intervalprolonging side effects of drugs in development. A major impediment to the widespread use of human iPSC-CM assays is the low throughput of the currently available electrophysiological tools. The patch-clamp technique allows for direct measurement of transmembrane voltage (V m ) to precisely quantify the morphology and duration of atrial and ventricular action potentials (APs). However, the patch-clamp technique is a labor-intensive technique, which involves highly skilled, experienced individuals. Recently, the utility of a genetically encoded voltage-sensitive fluorescent reporter (ArcLight) was described in human embryonic stem cell-derived CMs (6). This reporter generated robust fluorescent signals from single cells, allowing for moderate-high throughput analyses, although the temporal response was delayed relative to directly measured V m . While voltage-sensitive dyes such as di-4-ANNEPS and di-8-ANNEPS allow ...

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Section: Stability Of Di-4-anbdqbs Optical Ap Recordings and Photodynsupporting

confidence: 86%

Section: Methodsmentioning

confidence: 99%

A near-infrared fluorescent voltage-sensitive dye allows for moderate-throughput electrophysiological analyses of human induced pluripotent stem cell-derived cardiomyocytes

López-Izquierdo

Warren

Riedel

et al. 2014

American Journal of Physiology-Heart and Circulatory Physiology

Self Cite

View full text Add to dashboard Cite

show abstract

“…2) was comparable among hiPSC-CMs generated independently in difference labs, these two monolayer methods may be preferable for future studies using hiPSC-CMs as cardiac disease models. A recent report also characterized Ca handling of hiPSC-CMs generated by monolayer-based Matrigel Sandwich method [36]. However, this study only evaluated twitch Ca amplitude and morphology in response to different pacing rates, while SR function and Ca fluxes were not evaluated.…”

Section: Discussionmentioning

confidence: 99%

Comparable calcium handling of human iPSC-derived cardiomyocytes generated by multiple laboratories

Hwang

Kryshtal

Feaster

et al. 2015

Journal of Molecular and Cellular Cardiology

144

134

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Cardiomyocytes (CMs) derived from human induced pluripotent stem cells (hiPSCs) are being increasingly used to model human heart diseases. hiPSC-CMs generated by earlier aggregation-based methods (i.e., embryoid body) often lack functional sarcoplasmic reticulum (SR) Ca stores characteristic of mature mammalian CMs. Newer monolayer-based cardiac differentiation methods (i.e., Matrigel sandwich or small molecule-based differentiation) produce hiPSC-CMs of high purity and yield, but their Ca handling has not been comprehensively investigated. Here, we studied Ca handling and cytosolic Ca buffering properties of hiPSC-CMs generated independently from multiple hiPSC lines at Stanford University, Vanderbilt University and University of Wisconsin. hiPSC-CMs were cryopreserved at each university. Frozen aliquots were shipped, recovered from cryopreservation, plated at low density and compared 3-5 days after plating with acutely-isolated adult rabbit and mouse ventricular CMs. Although hiPSC-CM cell volume was significantly smaller, cell capacitance to cell volume ratio and cytoplasmic Ca buffering were not different from rabbit-CMs. hiPSC-CMs from all three laboratories exhibited robust L-type Ca currents, twitch Ca transients and caffeine-releasable SR Ca stores comparable to adult CMs. Ca transport by sarcoendoplasmic reticulum Ca ATPase (SERCA) and Na/Ca exchanger (NCX) was similar in all hiPSC-CM lines, but slower compared to rabbit-CMs. However, the relative contribution of SERCA and NCX to Ca transport of hiPSC-CMs was comparable to rabbit-CMs. Ca handling maturity of hiPSC-CMs increased from 15 to 21 days post-induction. We conclude that hiPSC-CMs generated independently from multiple iPSC lines using monolayer-based methods can be reproducibly recovered from cryopreservation and exhibit comparable and functional SR Ca handling.

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“…In their place, cardiac screening technologies are increasingly focusing on the use of either microelectrode arrays (MEAs) or optical mapping techniques to assess the electrophysiological output of cultured cardiomyocytes. MEAs permit recording of electrical field potentials from cardiac monolayers similar to whole heart analysis by electrocardiography (Clements and Thomas, 2014, Navarrete, Liang, 2013, Riedel et al, 2014), whereas the use of either genetically encoded or exogenously applied voltage-sensitive dyes enable real-time monitoring of action potential propagation through cultured cells (Kijlstra, Hu, 2015). Ca 2+ sensitive dyes have also been employed for analysis of cardiac function (Burridge, Thompson, 2011, Joshi-Mukherjee et al, 2013).…”

Section: Biomimetic Strategies For Human Cardiac Tissue Engineeringmentioning

confidence: 99%

Human iPSC-derived cardiomyocytes and tissue engineering strategies for disease modeling and drug screening

Smith

Macadangdang

Leung

et al. 2017

Biotechnology Advances

129

103

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Improved methodologies for modeling cardiac disease phenotypes and accurately screening the efficacy and toxicity of potential therapeutic compounds are actively being sought to advance drug development and improve disease modeling capabilities. To that end, much recent effort has been devoted to the development of novel engineered biomimetic cardiac tissue platforms that accurately recapitulate the structure and function of the human myocardium. Within the field of cardiac engineering, induced pluripotent stem cells (iPSCs) are an exciting tool that offer the potential to advance the current state of the art, as they are derived from somatic cells, enabling the development of personalized medical strategies and patient specific disease models. Here we review different aspects of iPSC-based cardiac engineering technologies. We highlight methods for producing iPSC-derived cardiomyocytes (iPSC-CMs) and discuss their application to compound efficacy/toxicity screening and in vitro modeling of prevalent cardiac diseases. Special attention is paid to the application of micro- and nano-engineering techniques for the development of novel iPSC-CM based platforms and their potential to advance current preclinical screening modalities.

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Functional and Pharmacological Analysis of Cardiomyocytes Differentiated from Human Peripheral Blood Mononuclear-Derived Pluripotent Stem Cells

Cited by 39 publications

References 47 publications

A near-infrared fluorescent voltage-sensitive dye allows for moderate-throughput electrophysiological analyses of human induced pluripotent stem cell-derived cardiomyocytes

A near-infrared fluorescent voltage-sensitive dye allows for moderate-throughput electrophysiological analyses of human induced pluripotent stem cell-derived cardiomyocytes

Comparable calcium handling of human iPSC-derived cardiomyocytes generated by multiple laboratories

Human iPSC-derived cardiomyocytes and tissue engineering strategies for disease modeling and drug screening

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