scholarly journals Cell culture conditions affect the ability of high content imaging assay to detect drug-induced changes in cellular parameters in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs)

2019 ◽  
Vol 6 ◽  
pp. 305-320
Author(s):  
Bharathi Balasubramanian ◽  
Vaclav Belak ◽  
Isha Verma ◽  
Yeva Prysiazhniuk ◽  
Frederick Sannajust ◽  
...  
Keyword(s):  
Cell Culture ◽  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Culture Conditions ◽  
Drug Induced ◽  
High Content Imaging ◽  
Induced Pluripotent ◽  
Induced Changes ◽  
Cell Culture Conditions

scholarly journals Nanofibrous Matrix of Defined Composition Sustains Human Induced Pluripotent Stem Cell Culture

2021 ◽  
Vol 4 (4) ◽  
pp. 3035-3040
Author(s):  
Sara Borrego-González ◽  
Berta de la Cerda ◽  
Francisco J. Díaz-Corrales ◽  
Aránzazu Díaz-Cuenca
Keyword(s):  
Cell Culture ◽  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Stem Cell Culture ◽  
Induced Pluripotent

Abstract 142: Modeling Drug-Induced Long QT Syndrome with Patient-Specific Induced Pluripotent Stem Cell-Derived Cardiomyocytes

2013 ◽  
Vol 113 (suppl_1) ◽  
Author(s):  
Francesca Stillitano ◽  
Ioannis Karakikes ◽  
Chi-wai Kong ◽  
Brett Martinelli ◽  
Ronald Li ◽  
...  
Keyword(s):  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Patient Specific ◽  
Dependent Manner ◽  
Long Qt ◽  
Drug Induced ◽  
Qt Syndrome ◽  
Induced Pluripotent ◽  
Dose Dependent

Long QT syndrome (LQTS) is characterized by prolonged cardiac repolarization time and increased risk of ventricular arrhythmia. LQTS can be either inherited or induced notably after drugs intake. Mutations in genes encoding cardiac ion channels have been reported to underlie inherited LQTS. In contrast, drug-induced LQTS (diLQTS) most frequently arises from altered function of the hERG channel; the risk of developing diLQTS varies largely between subjects and most people who have life-threatening diLQTS have no known genetic risk factors. We investigated whether the susceptibility to develop diLQTS observed in vivo can be recapitulated in vitro using patient-specific induced pluripotent stem cell (iPSC) technology. We collected skin fibroblasts from ten subjects who developed significant diLQTS after administration of Sotalol and/or Erythromycin. Ten other individuals who displayed no changes in QT interval after administration of the same drugs, were selected. iPSC were generated by retroviral delivery of Oct4, Sox2, Nanog and Klf4 in 17 of the 20 individuals. We report preliminary results obtained from iPSC-derived cardiomyocytes (iPSC-CMs) of two subjects. All experiments were performed in a blinded fashion without knowledge of the associated clinical phenotype. Cardiac differentiation of iPSC resulted in the generation of spontaneously beating embryoid bodies. iPSC-CMs showed positive staining for TNNT2, ACTN2 and Cx43. Gene expression analysis confirmed the expression of NKX2.5, MLC2v, MYH6 and MYH7, and of the relevant KCNH2 gene. The two lines had similar basal electrophysiological properties as assessed by measurements of action potential (AP) by patch-clamp technique and extracellular field potentials (FP) using micro-electrode array (MEA). E4031, a classical HERG blocker, significantly prolonged the FP duration (FPD) in a dose-dependent manner in both lines (EC50: 30.19 and 51.57 respectively). When both Sotalol and Erythromicin were used, FPD was prolonged in one of the two samples in a dose-dependent manner (EC50Sotalol: 100; EC50Erythr: 9.64) while drug response was blunted in the other cell line. This study suggests that patient-specific iPSC can be used to model the functional abnormalities observed in acquired diLQTS.

Time-course colony tracking analysis for evaluating induced pluripotent stem cell culture processes

2019 ◽  
Vol 128 (2) ◽  
pp. 209-217 ◽  
Author(s):  
Kei Yoshida ◽  
Mika Okada ◽  
Risako Nagasaka ◽  
Hiroto Sasaki ◽  
Mai Okada ◽  
...  
Keyword(s):  
Cell Culture ◽  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Time Course ◽  
Induced Pluripotent Stem Cell ◽  
Stem Cell Culture ◽  
Induced Pluripotent

scholarly journals Use of Human Induced Pluripotent Stem Cell–Derived Cardiomyocytes in Preclinical Cancer Drug Cardiotoxicity Testing: A Scientific Statement From the American Heart Association

2019 ◽  
Vol 125 (10) ◽  
Author(s):  
Gary Gintant ◽  
Paul Burridge ◽  
Lior Gepstein ◽  
Sian Harding ◽  
Todd Herron ◽  
...  
Keyword(s):  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Cardiac Injury ◽  
Induced Pluripotent Stem Cell ◽  
Heart Association ◽  
Cancer Drug ◽  
Great Promise ◽  
Drug Induced ◽  
Induced Pluripotent

It is now well recognized that many lifesaving oncology drugs may adversely affect the heart and cardiovascular system, including causing irreversible cardiac injury that can result in reduced quality of life. These effects, which may manifest in the short term or long term, are mechanistically not well understood. Research is hampered by the reliance on whole-animal models of cardiotoxicity that may fail to reflect the fundamental biology or cardiotoxic responses of the human myocardium. The emergence of human induced pluripotent stem cell–derived cardiomyocytes as an in vitro research tool holds great promise for understanding drug-induced cardiotoxicity of oncological drugs that may manifest as contractile and electrophysiological dysfunction, as well as structural abnormalities, making it possible to deliver novel drugs free from cardiac liabilities and guide personalized therapy. This article briefly reviews the challenges of cardio-oncology, the strengths and limitations of using human induced pluripotent stem cell–derived cardiomyocytes to represent clinical findings in the nonclinical research space, and future directions for their further use.

scholarly journals Blinded, Multicenter Evaluation of Drug-induced Changes in Contractility Using Human-induced Pluripotent Stem Cell-derived Cardiomyocytes

2020 ◽  
Vol 176 (1) ◽  
pp. 103-123 ◽  
Author(s):  
Umber Saleem ◽  
Berend J van Meer ◽  
Puspita A Katili ◽  
Nurul A N Mohd Yusof ◽  
Ingra Mannhardt ◽  
...  
Keyword(s):  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Signal To Noise Ratio ◽  
Safety Evaluation ◽  
Primary Source ◽  
Induced Pluripotent Stem Cell ◽  
Drug Induced ◽  
Inotropic Effects ◽  
Induced Pluripotent ◽  
Improving Accuracy

Abstract Animal models are 78% accurate in determining whether drugs will alter contractility of the human heart. To evaluate the suitability of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) for predictive safety pharmacology, we quantified changes in contractility, voltage, and/or Ca2+ handling in 2D monolayers or 3D engineered heart tissues (EHTs). Protocols were unified via a drug training set, allowing subsequent blinded multicenter evaluation of drugs with known positive, negative, or neutral inotropic effects. Accuracy ranged from 44% to 85% across the platform-cell configurations, indicating the need to refine test conditions. This was achieved by adopting approaches to reduce signal-to-noise ratio, reduce spontaneous beat rate to ≤ 1 Hz or enable chronic testing, improving accuracy to 85% for monolayers and 93% for EHTs. Contraction amplitude was a good predictor of negative inotropes across all the platform-cell configurations and of positive inotropes in the 3D EHTs. Although contraction- and relaxation-time provided confirmatory readouts forpositive inotropes in 3D EHTs, these parameters typically served as the primary source of predictivity in 2D. The reliance of these “secondary” parameters to inotropy in the 2D systems was not automatically intuitive and may be a quirk of hiPSC-CMs, hence require adaptations in interpreting the data from this model system. Of the platform-cell configurations, responses in EHTs aligned most closely to the free therapeutic plasma concentration. This study adds to the notion that hiPSC-CMs could add value to drug safety evaluation.

scholarly journals Stem cell culture conditions and stability: a joint workshop of the PluriMes Consortium and Pluripotent Stem Cell Platform

2019 ◽  
Vol 14 (3) ◽  
pp. 243-255 ◽  
Author(s):  
Glyn N Stacey ◽  
Peter W Andrews ◽  
Ivana Barbaric ◽  
Charlotta Boiers ◽  
Amit Chandra ◽  
...  
Keyword(s):  
Cell Culture ◽  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Culture Conditions ◽  
Stem Cell Culture ◽  
Joint Workshop ◽  
Cell Culture Conditions

scholarly journals A Novel Purification Method of Murine Embryonic Stem Cell– and Human-Induced Pluripotent Stem Cell–Derived Cardiomyocytes by Simple Manual Dissociation

2012 ◽  
Vol 17 (5) ◽  
pp. 683-691 ◽  
Author(s):  
Tadahiro Shinozawa ◽  
Hatsue Furukawa ◽  
Eimei Sato ◽  
Kenji Takami
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Pluripotent Stem Cell ◽  
Embryonic Stem ◽  
Induced Pluripotent Stem Cell ◽  
Drug Induced ◽  
Thin Glass ◽  
Murine Embryonic Stem Cell ◽  
Induced Pluripotent

Cardiomyocytes derived from embryonic stem cells (ES-CMs) and induced pluripotent stem cells (iPS-CMs) are useful for toxicity and pharmacology screening. In the present study, we found that cardiomyocyte-rich beating cell clusters (CCs) emerged from murine embryonic stem cell (mESC)–derived beating EBs and from human-induced pluripotent stem cell (hiPSC)–derived beating EBs dissociated by gentle pipetting with a thin glass pipette. The percentage of cardiac troponin T (cTnT)–positive cells in the beating CCs obtained from mESC-derived and hiPSC-derived beating EBs was higher (81.5% and 91.6%, respectively) than in beating-undissociated EBs (13.7% and 67.1%, respectively). For mESCs, the yield of cTnT-positive cells from beating CCs was estimated to be 1.6 times higher than that of beating EBs. The bromodeoxyuridine labeling index of mouse ES-CMs and human iPS-CMs in beating CCs was 1.5- and 3.2-fold, respectively, greater than those in beating EBs. To investigate the utility of the cells in toxicity assessment, we showed that doxorubicin, a cardiotoxic drug, induced myofilament disruption in cardiomyocytes isolated by this method. This simple method enables preparation of mouse ES-CMs and human iPS-CMs with better proliferative activity than beating EBs not dissociated by pipetting, and the cardiomyocytes are useful for drug-induced myocardial toxicity testing.

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