Pluripotent stem cells as a platform for cardiac arrhythmia drug screening

Current Treatment Options in Cardiovascular Medicine

Volumn 16, Issue 9, 2014, Pages

  Leyton Mange, Jordan S ^a   Milan, David J ^a  

^a MASSACHUSETTS GENERAL HOSPITAL   (United States)

Author keywords

Arrhythmia; Cardiac electrophysiology; Cardiomyocytes; Electrophysiology; Pluripotent; Screening; Stem cell

Indexed keywords

ACTION POTENTIAL; BIOASSAY; CARDIOTOXICITY; CATECHOLAMINERGIC POLYMORPHIC VENTRICULAR TACHYCARDIA; CELL DIFFERENTIATION; CONGESTIVE CARDIOMYOPATHY; CONTRACTILE IMPEDANCE ANALYSIS; DRUG SCREENING; ELECTRIC ACTIVITY; ELECTRODE; FLUORESCENCE IMAGING; FLUORESCENCE OPTICAL MAAPING; HEART ARRHYTHMIA; HEART ELECTROPHYSIOLOGY; HEART MUSCLE CELL; HEART MUSCLE CONTRACTILITY; HEART RIGHT VENTRICLE DYSPLASIA; HUMAN; HYPERTROPHIC CARDIOMYOPATHY; IMAGE ANALYSIS; IMPEDANCE; LEOPARD SYNDROME; LONG QT SYNDROME; MULTI ELECTRODE ARRAY; NONHUMAN; PHENOTYPE; PLURIPOTENT STEM CELL; REVIEW;

EID: 84904850251     PISSN: 10928464     EISSN: 15343189     Source Type: Journal    
DOI: 10.1007/s11936-014-0334-1     Document Type: Review

References (100)

1. Maltsev VA, Wobus AM, Rohwedel J, Bader M, Hescheler J. Cardiomyocytes differentiated in vitro from embryonic stem cells developmentally express cardiac-specific genes and ionic currents. Circ Res. 1994;75(2):233-44. (Pubitemid 24228315)
2. •• Kehat I, Kenyagin-Karsenti D, Snir M, Segev H, Amit M, Gepstein A, et al. Human embryonic stem cells can differentiate into myocytes with structural and functional properties of cardiomyocytes. J Clin Investig. 2001;108(3):407-14. doi:10.1172/JCI12131. Initial demonstration of human embryonic stem cells differentiated by the embryonic body method can form functional cardiomyocytes
3. Xu C, Police S, Rao N, Carpenter MK. Characterization and enrichment of cardiomyocytes derived from human embryonic stem cells. Circ Res. 2002;91(6):501-8.
4. •• Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006;126(4):663-76. doi:10.1016/j.cell.2006.07.024. Nobel-prize winning work from the Yamanaka lab demonstrating that mature somatic cells can be reprogrammed to a pluripotent state
5. •• Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, et al. Multiplex genome engineering using CRISPR/Cas systems. Science. 2013;339(6121):819-23. doi:10.1126/science.1231143. Independently, members of the Church and Zhang labs (see reference #6 below) demonstrated use of the CRISPR/Cas9 system in targeted mutagenesis in mammalian cells, marking a new age in genome editing technology
6. •• Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE, et al. RNA-guided human genome engineering via Cas9. Science. 2013;339(6121):823-6. doi:10.1126/science.1232033.
7. Musunuru K. Genome editing of human pluripotent stem cells to generate human cellular disease models. Dis Models Mech. 2013;6(4):896-904. doi:10.1242/dmm.012054.
8. •• Moretti A, Bellin M, Welling A, Jung CB, Lam JT, Bott-Flugel L, et al. Patient-specific induced pluripotent stem-cell models for long-QT syndrome. New Engl J Med. 2010;363(15):1397-409. doi:10.1056/NEJMoa0908679. Initial demonstration of an arrhythmogenic disease, in this case LQTI, modeled in human iPSC-derived cardiomyocytes
9. Itzhaki I, Maizels L, Huber I, Zwi-Dantsis L, Caspi O, Winterstern A, et al. Modelling the long QT syndrome with induced pluripotent stem cells. Nature. 2011;471(7337):225-9. doi:10.1038/nature09747.
10. Matsa E, Rajamohan D, Dick E, Young L, Mellor I, Staniforth A, et al. Drug evaluation in cardiomyocytes derived from human induced pluripotent stem cells carrying a long QT syndrome type 2 mutation. Eur Heart J. 2011;32(8):952-62. doi:10.1093/eurheartj/ehr073.
11. Yazawa M, Hsueh B, Jia X, Pasca AM, Bernstein JA, Hallmayer J, et al. Using induced pluripotent stem cells to investigate cardiac phenotypes in Timothy syndrome. Nature. 2011;471(7337):230-4. doi:10.1038/nature09855.
12. Davis RP, Casini S, van den Berg CW, Hoekstra M, Remme CA, Dambrot C, et al. Cardiomyocytes derived from pluripotent stem cells recapitulate electrophysiological characteristics of an overlap syndrome of cardiac sodium channel disease. Circulation. 2012;125(25):3079-91. doi:10.1161/CIRCULATIONAHA. 111.066092.
13. Itzhaki I, Maizels L, Huber I, Gepstein A, Arbel G, Caspi O, et al. Modeling of catecholaminergic polymorphic ventricular tachycardia with patient-specific human-induced pluripotent stem cells. J Am Coll Cardiol. 2012;60(11):990 -1000. doi:10.1016/j.jacc.2012.02.066.
14. Kim C, Wong J, Wen J, Wang S, Wang C, Spiering S, et al. Studying arrhythmogenic right ventricular dysplasia with patient-specific iPSCs. Nature. 2013;494(7435):105-10. doi:10.1038/nature11799.
15. Ma D, Wei H, Lu J, Ho S, Zhang G, Sun X, et al. Generation of patient-specific induced pluripotent stem cell-derived cardiomyocytes as a cellular model of arrhythmogenic right ventricular cardiomyopathy. Eur Heart J. 2013;34(15):1122-33. doi:10.1093/eurheartj/ehs226.
16. Roden DM, Hong CC. Stem cell-derived cardiomyocytes as a tool for studying proarrhythmia: a better canary in the coal mine? Circulation. 2013;127(16):1641-3. doi:10.1161/CIRCULATIONAHA.113.002127.
17. Sun N, Yazawa M, Liu J, Han L, Sanchez-Freire V, Abilez OJ, et al. Patient-specific induced pluripotent stem cells as a model for familial dilated cardiomyopathy. Sci Transl Med. 2012;4(130):130ra47. doi:10.1126/scitranslmed. 3003552.
18. Lan F, Lee AS, Liang P, Sanchez-Freire V, Nguyen PK, Wang L, et al. Abnormal calcium handling properties underlie familial hypertrophic cardiomyopathy pathology in patient-specific induced pluripotent stem cells. Cell Stem Cell. 2013;12(1):101-13. doi:10.1016/j.stem.2012.10.010.
19. Roden DM. Drug-induced prolongation of the QT interval. New Engl J Med. 2004;350(10):1013-22. doi:10.1056/NEJMra032426. (Pubitemid 38269278)
20. •• Liang P, Lan F, Lee AS, Gong T, Sanchez-Freire V, Wang Y, et al. Drug screening using a library of human induced pluripotent stem cell-derived cardiomyocytes reveals disease-specific patterns of cardiotoxicity. Circulation. 2013;127(16):1677-91. doi:10.1161/CIRCULATIONAHA.113.001883. Using patch clamp electrophysiology, the Wu laboratory at Stanford compared multiple human iPSC-CMs: those derived from patients with the LQT1 syndrome, hypertrophic cardiomyopathy (β-MHC mutation), and dilated cardiomyopathy (TNNT2 mutation), and found drug toxicity effects (i.e., EADs in LQT1 cells) in increased frequency in diseased lines compared to lines created from sibling controls or ES cells
21. Khan JM, Lyon AR, Harding SE. The case for induced pluripotent stem cell-derived cardiomyocytes in pharmacological screening. Br J Pharmacol. 2013;169(2):304-17. doi:10.1111/j.1476-5381.2012.02118.x.
22. Mercola M, Colas A, Willems E. Induced pluripotent stem cells in cardiovascular drug discovery. Circ Res. 2013;112(3):534-48. doi:10.1161/CIRCRESAHA.111.250266.
23. •• Zhang J, Klos M, Wilson GF, Herman AM, Lian X, Raval KK, et al. Extracellular matrix promotes highly efficient cardiac differentiation of human pluripotent stem cells: the matrix sandwich method. Circ Res. 2012;111(9):1125- 36. doi:10.1161/CIRCRESAHA.112.273144. The "matrix-sandwich" method is a highly preferred method of inducing cardiac differentaition in a monolayer format in hPSCs, with efficiencies as high as 9 80%
24. Lian X, Hsiao C, Wilson G, Zhu K, Hazeltine LB, Azarin SM, et al. Robust cardiomyocyte differentiation from human pluripotent stem cells via temporal modulation of canonical Wnt signaling. Proc Natl Acad Sci. 2012;109(27):E1848- 57. doi:10.1073/pnas.1200250109.
25. Chen G, Gulbranson DR, Hou Z, Bolin JM, Ruotti V, Probasco MD, et al. Chemically defined conditions for human iPSC derivation and culture. Nat Methods. 2011;8(5):424-9. doi:10.1038/nmeth.1593.
26. Magyar J, Iost N, Kortvely A, Banyasz T, Virag L, Szigligeti P, et al. Effects of endothelin-1 on calcium and potassium currents in undiseased human ventricular myocytes. Pflugers Arch. 2000;441(1):144-9. (Pubitemid 32146969)
27. • Hoekstra M, Mummery CL, Wilde AAM, Bezzina CR, Verkerk AO. Induced pluripotent stem-cell-derived cardiomyocytes asmodels for cardiac arrhythmias. Front Physiol. 2012;3. doi:10.3389/fphys.2012.00346. One of the most comprehensive reviews of utilizing hPSCCMs as models for arrhythmia. The authors review, individually, the present body of work of iPSC models for various LQT syndromes and CPVT. Authors also review evidence for the major cardiac currents and their respective properties in hPSC-CMs.
28. Peng S, Lacerda AE, Kirsch GE, Brown AM, Bruening- Wright A. The action potential and comparative pharmacology of stem cell-derived human cardiomyocytes. J Pharmacol Toxicol Meth. 2010;61(3):277-86. doi:10.1016/j.vascn.2010.01.014.
29. • Mummery C, Ward-van Oostwaard D, Doevendans P, Spijker R, van den Brink S, Hassink R, et al. Differentiation of human embryonic stem cells to cardiomyocytes: role of coculture with visceral endoderm-like cells. Circulation. 2003;107(21):2733-40. doi:10.1161/01.CIR.0000068356.38592.68. The visceral endoderm coculture method of cardiomyocyte differentation from hPSCs
30. Robertson C, Tran DD, George SC. Concise review: maturation phases of human pluripotent stem cell-derived cardiomyocytes. Stem Cells. 2013;31(5):829 -37. doi:10.1002/stem.1331.
31. Bellin M, Casini S, Davis RP, D'Aniello C, Haas J, Wardvan Oostwaard D, et al. Isogenic human pluripotent stem cell pairs reveal the role of a KCNH2 mutation in long-QT syndrome. EMBO J. 2013;32(24):3161-75. doi:10.1038/emboj. 2013.240.
32. • Knollmann BC. Induced pluripotent stem cell-derived cardiomyocytes: boutique science or valuable arrhythmia model? Circ Res. 2013;112(6):969-76; discussion 76. doi:10.1161/CIRCRESAHA.112.300567. It has increasingly been recognized that genetic differences between iPSC and ESC lines may underlie variability in their respective phenotypes and drug responses. These authors generated isogenic control lines via genome editing by correcting a KCNH2 mutation in a LQT2 hiPSC line and introducing it into a WT hESC control line. By making comparisons within the same genetic background, such isogenic control strategies may emerge as a preferred method to compare phenotypic effects of mutations.
33. Zhang Q, Jiang J, Han P, Yuan Q, Zhang J, Zhang X, et al. Direct differentiation of atrial and ventricular myocytes from human embryonic stem cells by alternating retinoid signals. Cell Res. 2011;21(4):579-87. doi:10.1038/cr.2010.163.
34. • Ma J, Guo L, Fiene SJ, Anson BD, Thomson JA, Kamp TJ, et al. High purity human-induced pluripotent stem cell-derived cardiomyocytes: electrophysiological properties of action potentials and ionic currents. Am J Physiol Heart Circ Physiol. 2011;301(5):H2006-17. doi:10.1152/ajpheart.00694. 2011. The January group rigorously evaluated various cardiac currents in cells from one single iPSC line from Cellular Dynamics, Madison, WI
35. Satin J, Kehat I, Caspi O, Huber I, Arbel G, Itzhaki I, et al. Mechanism of spontaneous excitability in human embryonic stem-cell-derived cardiomyocytes. J Physiol. 2004;559(Pt 2):479-96. doi:10.1113/jphysiol.2004.068213. (Pubitemid 39263813)
36. Wang K, Terrenoire C, Sampson KJ, Iyer V, Osteen JD, Lu J, et al. Biophysical properties of slow potassium channels in human embryonic stem-cell-derived cardiomyocytes implicate subunit stoichiometry. J Physiol. 2011;589(Pt 24):6093-104. doi:10.1113/jphysiol.2011.220863.
37. Bett GC, Kaplan AD, Lis A, Cimato TR, Tzanakakis ES, Zhou Q, et al. Electronic "expression" of the inward rectifier in cardiocytes derived from human-induced pluripotent stem cells. Heart Rhythm. 2013;10(12):1903-10. doi:10.1016/j.hrthm.2013.09.061.
38. Itzhaki I, Rapoport S, Huber I, Mizrahi I, Zwi-Dantsis L, Arbel G, et al. Calcium handling in human induced pluripotent stem-cell -derived cardiomyocytes. PLoS One. 2011;6(4):e18037. doi:10.1371/journal.pone.0018037.
39. Norstrom A, Akesson K, Hardarson T, Hamberger L, Bjorquist P, Sartipy P. Molecular and pharmacological properties of human embryonic stem cell-derived cardiomyocytes. Exp Biol Med (Maywood). 2006;231(11):1753 -62.
40. Guo L, Coyle L, Abrams RM, Kemper R, Chiao ET, Kolaja KL. Refining the Human iPSC-Cardiomyocyte Arrhythmic Risk Assessment Model. Toxicol Sci. 2013;136(2):581-94. doi:10.1093/toxsci/kft205.
41. Sheng X, Reppel M, Nguemo F, Mohammad FI, Kuzmenkin A, Hescheler J, et al. Human pluripotent stem cell-derived cardiomyocytes: response to TTX and lidocain reveals strong cell to cell variability. PLoS One. 2012;7(9):e45963. doi:10.1371/journal.pone.0045963.
42. Egashira T, Yuasa S, Suzuki T, Aizawa Y, Yamakawa H, Matsuhashi T, et al. Disease characterization using LQTS-specific induced pluripotent stem cells. Cardiovasc Res. 2012;95(4):419 -29. doi:10.1093/cvr/cvs206.
43. Lahti AL, Kujala VJ, Chapman H, Koivisto AP, Pekkanen-Mattila M, Kerkela E, et al. Model for long QT syndrome type 2 using human iPS cells demonstrates arrhythmogenic characteristics in cell culture. Dis Models Mech. 2012;5(2):220-30. doi:10.1242/dmm.008409.
44. Ma D, Wei H, Zhao Y, Lu J, Li G, Sahib NB, et al. Modeling type 3 long QT syndrome with cardiomyocytes derived from patient-specific induced pluripotent stem cells. Int J Cardiol. 2013;168(6):5277- 86. doi:10.1016/j.ijcard.2013.08. 015.
45. Fatima A, Kaifeng S, Dittmann S, Xu G, Gupta MK, Linke M, et al. The Disease-Specific Phenotype in Cardiomyocytes Derived from Induced Pluripotent Stem Cells of Two Long QT Syndrome Type 3 Patients. PLoS One. 2013;8(12):e83005. doi:10.1371/journal.pone.0083005.
46. Terrenoire C, Wang K, Tung KW, Chung WK, Pass RH, Lu JT, et al. Induced pluripotent stem cells used to reveal drug actions in a long QT syndrome family with complex genetics. J Gen Physiol. 2013;141(1):61-72. doi:10.1085/jgp. 201210899.
47. Jonsson MK, Vos MA, Mirams GR, Duker G, Sartipy P, de Boer TP, et al. Application of human stem cell-derived cardiomyocytes in safety pharmacology requires caution beyond hERG. J Mol Cell Cardiol. 2012;52(5):998-1008. doi:10.1016/j.yjmcc.2012.02.002.
48. Jung CB, Moretti A, Mederos y Schnitzler M, Iop L, Storch U, Bellin M, et al. Dantrolene rescues arrhythmogenic RYR2 defect in a patient-specific stem cell model of catecholaminergic polymorphic ventricular tachycardia. EMBO Mol Med. 2012;4(3):180-91. doi:10.1002/emmm.201100194.
49. Kujala K, Paavola J, Lahti A, Larsson K, Pekkanen- Mattila M, Viitasalo M, et al. Cell model of catecholaminergic polymorphic ventricular tachycardia reveals early and delayed afterdepolarizations. PLoS One. 2012;7(9):e44660. doi:10.1371/journal.pone.0044660.
50. Zhang XH, Haviland S, Wei H, Saric T, Fatima A, Hescheler J, et al. Ca2+ signaling in human induced pluripotent stem cell-derived cardiomyocytes (iPSCM) from normal and catecholaminergic polymorphic ventricular tachycardia (CPVT)-afflicted subjects. Cell Calcium. 2013;54(2):57 -70. doi:10.1016/j.ceca. 2013.04.004.
51. Novak A, Barad L, Zeevi-Levin N, Shick R, Shtrichman R, Lorber A, et al. Cardiomyocytes generated from CPVTD307H patients are arrhythmogenic in response to beta-adrenergic stimulation. J Cell Mol Med. 2012;16(3):468-82. doi:10.1111/j.1582-4934.2011.01476.x.
52. Satin J, Itzhaki I, Rapoport S, Schroder EA, Izu L, Arbel G, et al. Calcium handling in human embryonic stem cell-derived cardiomyocytes. Stem Cells. 2008;26(8):1961-72. doi:10.1634/stemcells.2007-0591.
53. Caspi O, Huber I, Gepstein A, Arbel G, Maizels L, Boulos M, et al. Modeling of arrhythmogenic right ventricular cardiomyopathy with human induced pluripotent stem cells. Circ Cardiovasc Genet. 2013;6(6):557-68. doi:10.1161/CIRCGENETICS.113.000188.
54. Huang HP, Chen PH, Hwu WL, Chuang CY, Chien YH, Stone L, et al. Human Pompe disease-induced pluripotent stem cells for pathogenesis modeling, drug testing and disease marker identification. Hum Mol Genet. 2011;20(24):4851-64. doi:10.1093/hmg/ddr424.
55. Zwi L, Caspi O, Arbel G, Huber I, Gepstein A, Park IH, et al. Cardiomyocyte differentiation of human induced pluripotent stem cells. Circulation. 2009;120(15):1513-23. doi:10.1161/CIRCULATIONAHA.109.868885.
56. Mummery CL, Zhang J, Ng ES, Elliott DA, Elefanty AG, Kamp TJ. Differentiation of human embryonic stem cells and induced pluripotent stem cells to cardiomyocytes: a methods overview. Circ Res. 2012;111(3):344-58. doi:10.1161/CIRCRESAHA.110.227512.
57. Burridge PW, Thompson S, Millrod MA, Weinberg S, Yuan X, Peters A, et al. A universal system for highly efficient cardiac differentiation of human induced pluripotent stem cells that eliminates interline variability. PLoS One. 2011;6(4):e18293. doi:10.1371/journal.pone.0018293.
58. Xu C, Police S, Hassanipour M, Li Y, Chen Y, Priest C, et al. Efficient generation and cryopreservation of cardiomyocytes derived from human embryonic stem cells. Regen Med. 2011;6(1):53-66. doi:10.2217/rme.10.91.
59. Paige SL, Osugi T, Afanasiev OK, Pabon L, Reinecke H, Murry CE. Endogenous Wnt/beta-catenin signaling is required for cardiac differentiation in human embryonic stem cells. PLoS One. 2010;5(6):e11134. doi:10.1371/journal. pone.0011134.
60. Cao N, Liu Z, Chen Z, Wang J, Chen T, Zhao X, et al. Ascorbic acid enhances the cardiac differentiation of induced pluripotent stem cells through promoting the proliferation of cardiac progenitor cells. Cell Res. 2012;22(1):219-36. doi:10.1038/cr.2011.195.
61. Chetty S, Pagliuca FW, Honore C, Kweudjeu A, Rezania A, Melton DA. A simple tool to improve pluripotent stem cell differentiation. Nat Methods. 2013;10(6):553-6. doi:10.1038/nmeth.2442.
62. Xu C. Differentiation and enrichment of cardiomyocytes from human pluripotent stem cells. J Mol Cell Cardiol. 2012;52(6):1203-12. doi:10.1016/j.yjmcc.2012.03.012.
63. Bizy A, Guerrero-Serna G, Hu B, Ponce-Balbuena D, Willis BC, Zarzoso M, et al. Myosin light chain 2- based selection of human iPSC-derived early ventricular cardiac myocytes. Stem Cell Res. 2013;11(3):1335-47. doi:10.1016/j.scr.2013.09.003.
64. Elliott DA, Braam SR, Koutsis K, Ng ES, Jenny R, Lagerqvist EL, et al. NKX2-5(eGFP/w) hESCs for isolation of human cardiac progenitors and cardiomyocytes. Nat Methods. 2011;8(12):1037-40. doi:10.1038/nmeth.1740.
65. • Kita-Matsuo H, Barcova M, Prigozhina N, Salomonis N, Wei K, Jacot JG, et al. Lentiviral vectors and protocols for creation of stable hESC lines for fluorescent tracking and drug resistance selection of cardiomyocytes. PLoS One. 2009;4(4):e5046. doi:10.1371/journal.pone.0005046. Enrichment of cardiomyocyte populations from heterogeneous differentiated preparations of hPSCs can be done by various methods. Transgenic α-MHC promoter driven selectional markers is a common preferred method across many groups
66. Tohyama S, Hattori F, Sano M, Hishiki T, Nagahata Y, Matsuura T, et al. Distinct metabolic flow enables largescale purification ofmouse and human pluripotent stem cell-derived cardiomyocytes. Cell Stem Cell. 2013;12(1):127-37. doi:10.1016/j.stem.2012.09.013.
67. Hattori F, Chen H, Yamashita H, Tohyama S, Satoh YS, Yuasa S, et al. Nongenetic method for purifying stem cell-derived cardiomyocytes. Nat Methods. 2010;7(1):61-6. doi:10.1038/nmeth.1403.
68. Dubois NC, Craft AM, Sharma P, Elliott DA, Stanley EG, Elefanty AG, et al. SIRPA is a specific cell-surface marker for isolating cardiomyocytes derived from human pluripotent stem cells. Nat Biotechnol. 2011;29(11):1011- 8. doi:10.1038/nbt.2005.
69. Uosaki H, Fukushima H, Takeuchi A, Matsuoka S, Nakatsuji N, Yamanaka S, et al. Efficient and scalable purification of cardiomyocytes from human embryonic and induced pluripotent stem cells by VCAM1 surface expression. PLoS One. 2011;6(8):e23657. doi:10.1371/journal.pone.0023657.
70. Nunes SS, Miklas JW, Liu J, Aschar-Sobbi R, Xiao Y, Zhang B, et al. Biowire: a platform for maturation of human pluripotent stem cell-derived cardiomyocytes. Nat Methods. 2013;10(8):781-7. doi:10.1038/nmeth.2524.
71. Lieu DK, Fu JD, Chiamvimonvat N, Tung KC, McNerney GP, Huser T, et al. Mechanism-based facilitated maturation of human pluripotent stem cell-derived cardiomyocytes. Circ Arrhythmia Electrophysiol. 2013;6(1):191-201. doi:10.1161/CIRCEP.111.973420.
72. •• Lee P, Klos M, Bollensdorff C, Hou L, Ewart P, Kamp TJ, et al. Simultaneous voltage and calcium mapping of genetically purified human induced pluripotent stem cell-derived cardiac myocyte monolayers. Circ Res. 2012;110(12):1556-63. doi:10.1161/CIRCRESAHA.111.262535. Fluorescent indicators of voltage and calcium dynamics may emerge as a preferred, non-invasive method for hPSC-CM phenotyping, as undertaken in cell monolayers in this study by Lee and colleagues
73. Mitcheson JS, Hancox JC, Levi AJ. Cultured adult cardiac myocytes: future applications, culture methods, morphological and electrophysiological properties. Cardiovasc Res. 1998;39(2):280-300. (Pubitemid 28346525)
74. Louch WE, Sheehan KA, Wolska BM. Methods in cardiomyocyte isolation, culture, and gene transfer. J Mol Cell Cardiol. 2011;51(3):288-98. doi:10.1016/j.yjmcc.2011.06.012.
75. Kim C, Majdi M, Xia P, Wei KA, Talantova M, Spiering S, et al. Non-cardiomyocytes influence the electrophysiological maturation of human embryonic stem cell-derived cardiomyocytes during differentiation. Stem Cell Dev. 2010;19(6):783-95. doi:10.1089/scd.2009.0349.
76. Uesugi M, Ojima A, Taniguchi T, Miyamoto N, Sawada K. Low-density plating is sufficient to induce cardiac hypertrophy and electrical remodeling in highly purified human iPS cell-derived cardiomyocytes. J Pharmacol Toxicol Meth. 2013. doi:10.1016/j.vascn.2013.11.002.
77. Neher E, Sakmann B. Single-channel currents recorded from membrane of denervated frog muscle fibres. Nature. 1976;260(5554):799-802.
78. Milligan CJ, Moller C. Automated planar patchclamp. Methods Mol Biol. 2013;998:171-87. doi:10.1007/978-1-62703-351-0-13.
79. Stoelzle S, Obergrussberger A, Bruggemann A, Haarmann C, George M, Kettenhofen R, et al. State-of-the-Art Automated Patch Clamp Devices: Heat Activation, Action Potentials, and High Throughput in Ion Channel Screening. Front Pharmacol. 2011;2:76. doi:10.3389/fphar.2011.00076.
80. Yajuan X, Xin L, Zhiyuan L. A comparison of the performance and application differences between manual and automated patch-clamp techniques. Curr Chem Genom. 2012;6:87-92. doi:10.2174/1875397301206010087.
81. Harris K, Aylott M, Cui Y, Louttit JB, McMahon NC, Sridhar A. Comparison of electrophysiological data from human-induced pluripotent stem cell-derived cardiomyocytes to functional preclinical safety assays. Toxicol Sci. 2013;134(2):412-26. doi:10.1093/toxsci/kft113.
82. Igelmund P, Fleischmann BK, Fischer IR, Soest J, Gryshchenko O, Bohm-Pinger MM, et al. Action potential propagation failures in long-term recordings from embryonic stem cell-derived cardiomyocytes in tissue culture. Pflugers Arch. 1999;437(5):669-79. (Pubitemid 29161922)
83. Caspi O, Itzhaki I, Kehat I, Gepstein A, Arbel G, Huber I, et al. In vitro electrophysiological drug testing using human embryonic stem-cell-derived cardiomyocytes. Stem Cell Dev. 2009;18(1):161-72. doi:10.1089/scd.2007.0280.
84. Mandel Y, Weissman A, Schick R, Barad L, Novak A, Meiry G, et al. Human embryonic and induced pluripotent stem cell-derived cardiomyocytes exhibit beat rate variability and power-law behavior. Circulation. 2012;125(7):883-93. doi:10.1161/CIRCULATIONAHA.111.045146.
85. • Herron TJ, Lee P, Jalife J. Optical imaging of voltage and calcium in cardiac cells & tissues. Circ Res. 2012;110(4):609-23. doi:10.1161/CIRCRESAHA.111.247494. A comprehensive and up-to-date review of optical mapping of voltage and calcium signals using small molecule dyes in cardiac tissue and cells
86. Schaffer P, Ahammer H, Muller W, Koidl B, Windisch H. Di-4-ANEPPS causes photodynamic damage to isolated cardiomyocytes. Pflugers Arch. 1994;426(6):548-51. (Pubitemid 24131545)
87. Hardy ME, Lawrence CL, Standen NB, Rodrigo GC. Can optical recordings of membrane potential be used to screen for drug-induced action potential prolongation in single cardiac myocytes? J Pharmacol Toxicol Meth. 2006;54(2):173-82. doi:10.1016/j.vascn.2006.02.013. (Pubitemid 44382686)
88. Liau B, Zhang D, Bursac N. Functional cardiac tissue engineering. Regen Med. 2012;7(2):187-206. doi:10.2217/rme.11.122.
89. Sirenko O, Crittenden C, Callamaras N, Hesley J, Chen YW, Funes C, et al. Multiparameter in vitro assessment of compound effects on cardiomyocyte physiology using iPSC cells. J Biomol Screen. 2013;18(1):39-53. doi:10.1177/1087057112457590.
90. Cerignoli F, Charlot D, Whittaker R, Ingermanson R, Gehalot P, Savchenko A, et al. High throughput measurement of Ca(2)(+) dynamics for drug risk assessment in human stem cell-derived cardiomyocytes by kinetic image cytometry. J Pharmacol Toxicol Meth. 2012;66(3):246-56. doi:10.1016/j.vascn.2012.08.167.
91. Jin L, Han Z, Platisa J, Wooltorton JR, Cohen LB, Pieribone VA. Single action potentials and subthreshold electrical events imaged in neurons with a fluorescent protein voltage probe. Neuron. 2012;75(5):779-85. doi:10.1016/j.neuron.2012.06.040.
92. Cao G, Platisa J, Pieribone VA, Raccuglia D, Kunst M, Nitabach MN. Genetically targeted optical electrophysiology in intact neural circuits. Cell. 2013;154(4):904-13. doi:10.1016/j.cell.2013.07.027.
93. Kralj JM, Douglass AD, Hochbaum DR, Maclaurin D, Cohen AE. Optical recording of action potentials in mammalian neurons using a microbial rhodopsin. Nat Methods. 2012;9(1):90-5. doi:10.1038/nmeth.1782.
94. Tsutsui H, Jinno Y, Tomita A, Niino Y, Yamada Y, Mikoshiba K, et al. Improved detection of electrical activity with a voltage probe based on a voltage-sensing phosphatase. J Physiol. 2013;591(Pt 18):4427 -37. doi:10.1113/jphysiol.2013.257048.
95. Zhao Y, Araki S, Wu J, Teramoto T, Chang YF, Nakano M, et al. An expanded palette of genetically encoded Ca(2)(+) indicators. Science. 2011;333(6051):1888-91. doi:10.1126/science.1208592.
96. Leyton-Mange JS, Mills RW, Macri VS, Jang MY, Butte FN, Ellinor PT, et al. Rapid Cellular Phenotyping of Human Pluripotent Stem Cell-Derived Cardiomyocytes using a Genetically Encoded Fluorescent Voltage Sensor. Stem Cell Reports. 2014;2(2):163-70. doi:10.1016/j.stemcr.2014.01.003.
97. Sheehy SP, Pasqualini F, Grosberg A, Park SJ, Aratyn-Schaus Y, Parker KK. Quality metrics for stem cell-derived cardiac myocytes. Stem Cell Reports. 2014;2(3):282-94. doi:10.1016/j.stemcr.2014.01.015.
98. Chi KR. Revolution dawning in cardiotoxicity testing. Nat Rev Drug Discov. 2013;12(8):565-7. doi:10.1038/nrd4083.
99. Mehta A, Sequiera GL, Ramachandra CJ, Sudibyo Y, Chung Y, Jingwei S, et al. Re-trafficking of HERG Reverses Long QT Syndrome 2 Phenotype in Human iPS-derived Cardiomyocytes. Cardiovasc Res. 2014. doi:10.1093/cvr/cvu060.
100. Fatima A, Xu G, Shao K, Papadopoulos S, Lehmann M, Arnaiz-Cot JJ, et al. In vitro modeling of ryanodine receptor 2 dysfunction using human induced pluripotent stem cells. Cell Physiol Biochem: Int J Exp Cell Physiol Biochem Pharmacol. 2011;28(4):579-92. doi:10.1159/000335753.