Tissue-engineered Cardiac Patch for Advanced Functional Maturation of Human ESC-derived Cardiomyocytes

Overview

Journal Biomaterials

Specialty Biomedical Engineering

Date 2013 May 7

PMID 23642535

Citations 263

Authors

Donghui Zhang

Ilya Y Shadrin

Jason Lam

Hai-Qian Xian

H Ralph Snodgrass

Nenad Bursac

Affiliations

Soon will be listed here.

Abstract

Human embryonic stem cell-derived cardiomyocytes (hESC-CMs) provide a promising source for cell therapy and drug screening. Several high-yield protocols exist for hESC-CM production; however, methods to significantly advance hESC-CM maturation are still lacking. Building on our previous experience with mouse ESC-CMs, we investigated the effects of 3-dimensional (3D) tissue-engineered culture environment and cardiomyocyte purity on structural and functional maturation of hESC-CMs. 2D monolayer and 3D fibrin-based cardiac patch cultures were generated using dissociated cells from differentiated Hes2 embryoid bodies containing varying percentage (48-90%) of CD172a (SIRPA)-positive cardiomyocytes. hESC-CMs within the patch were aligned uniformly by locally controlling the direction of passive tension. Compared to hESC-CMs in age (2 weeks) and purity (48-65%) matched 2D monolayers, hESC-CMs in 3D patches exhibited significantly higher conduction velocities (CVs), longer sarcomeres (2.09 ± 0.02 vs. 1.77 ± 0.01 μm), and enhanced expression of genes involved in cardiac contractile function, including cTnT, αMHC, CASQ2 and SERCA2. The CVs in cardiac patches increased with cardiomyocyte purity, reaching 25.1 cm/s in patches constructed with 90% hESC-CMs. Maximum contractile force amplitudes and active stresses of cardiac patches averaged to 3.0 ± 1.1 mN and 11.8 ± 4.5 mN/mm(2), respectively. Moreover, contractile force per input cardiomyocyte averaged to 5.7 ± 1.1 nN/cell and showed a negative correlation with hESC-CM purity. Finally, patches exhibited significant positive inotropy with isoproterenol administration (1.7 ± 0.3-fold force increase, EC50 = 95.1 nm). These results demonstrate highly advanced levels of hESC-CM maturation after 2 weeks of 3D cardiac patch culture and carry important implications for future drug development and cell therapy studies.

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References

He J, Ma Y, Lee Y, Thomson J, Kamp T . Human embryonic stem cells develop into multiple types of cardiac myocytes: action potential characterization. Circ Res. 2003; 93(1):32-9. DOI: 10.1161/01.RES.0000080317.92718.99. View

Dolnikov K, Shilkrut M, Zeevi-Levin N, Gerecht-Nir S, Amit M, Danon A . Functional properties of human embryonic stem cell-derived cardiomyocytes: intracellular Ca2+ handling and the role of sarcoplasmic reticulum in the contraction. Stem Cells. 2005; 24(2):236-45. DOI: 10.1634/stemcells.2005-0036. View

Pillekamp F, Haustein M, Khalil M, Emmelheinz M, Nazzal R, Adelmann R . Contractile properties of early human embryonic stem cell-derived cardiomyocytes: beta-adrenergic stimulation induces positive chronotropy and lusitropy but not inotropy. Stem Cells Dev. 2012; 21(12):2111-21. DOI: 10.1089/scd.2011.0312. View

Pedrotty D, Klinger R, Kirkton R, Bursac N . Cardiac fibroblast paracrine factors alter impulse conduction and ion channel expression of neonatal rat cardiomyocytes. Cardiovasc Res. 2009; 83(4):688-97. PMC: 2725777. DOI: 10.1093/cvr/cvp164. View

Lee P, Klos M, Bollensdorff C, Hou L, Ewart P, Kamp T . Simultaneous voltage and calcium mapping of genetically purified human induced pluripotent stem cell-derived cardiac myocyte monolayers. Circ Res. 2012; 110(12):1556-63. PMC: 3423450. DOI: 10.1161/CIRCRESAHA.111.262535. View

Schaaf S, Shibamiya A, Mewe M, Eder A, Stohr A, Hirt M . Human engineered heart tissue as a versatile tool in basic research and preclinical toxicology. PLoS One. 2011; 6(10):e26397. PMC: 3197640. DOI: 10.1371/journal.pone.0026397. View

Reiser P, Portman M, Ning X, Schomisch Moravec C . Human cardiac myosin heavy chain isoforms in fetal and failing adult atria and ventricles. Am J Physiol Heart Circ Physiol. 2001; 280(4):H1814-20. DOI: 10.1152/ajpheart.2001.280.4.H1814. View

Tulloch N, Muskheli V, Razumova M, Korte F, Regnier M, Hauch K . Growth of engineered human myocardium with mechanical loading and vascular coculture. Circ Res. 2011; 109(1):47-59. PMC: 3140796. DOI: 10.1161/CIRCRESAHA.110.237206. View

Itzhaki I, Schiller J, Beyar R, Satin J, Gepstein L . Calcium handling in embryonic stem cell-derived cardiac myocytes: of mice and men. Ann N Y Acad Sci. 2006; 1080:207-15. DOI: 10.1196/annals.1380.017. View

10.

Binah O, Dolnikov K, Sadan O, Shilkrut M, Zeevi-Levin N, Amit M . Functional and developmental properties of human embryonic stem cells-derived cardiomyocytes. J Electrocardiol. 2007; 40(6 Suppl):S192-6. DOI: 10.1016/j.jelectrocard.2007.05.035. View

11.

Bian W, Liau B, Badie N, Bursac N . Mesoscopic hydrogel molding to control the 3D geometry of bioartificial muscle tissues. Nat Protoc. 2009; 4(10):1522-34. PMC: 2924624. DOI: 10.1038/nprot.2009.155. View

12.

Stevens K, Kreutziger K, Dupras S, Korte F, Regnier M, Muskheli V . Physiological function and transplantation of scaffold-free and vascularized human cardiac muscle tissue. Proc Natl Acad Sci U S A. 2009; 106(39):16568-73. PMC: 2746126. DOI: 10.1073/pnas.0908381106. View

13.

OHara T, Virag L, Varro A, Rudy Y . Simulation of the undiseased human cardiac ventricular action potential: model formulation and experimental validation. PLoS Comput Biol. 2011; 7(5):e1002061. PMC: 3102752. DOI: 10.1371/journal.pcbi.1002061. View

14.

Brandenburger M, Wenzel J, Bogdan R, Richardt D, Nguemo F, Reppel M . Organotypic slice culture from human adult ventricular myocardium. Cardiovasc Res. 2011; 93(1):50-9. DOI: 10.1093/cvr/cvr259. View

15.

Tohyama S, Hattori F, Sano M, Hishiki T, Nagahata Y, Matsuura T . Distinct metabolic flow enables large-scale purification of mouse and human pluripotent stem cell-derived cardiomyocytes. Cell Stem Cell. 2012; 12(1):127-37. DOI: 10.1016/j.stem.2012.09.013. View

16.

Flesch M, Schwinger R, SCHIFFER F, Frank K, Sudkamp M, Kuhn-Regnier F . Evidence for functional relevance of an enhanced expression of the Na(+)-Ca2+ exchanger in failing human myocardium. Circulation. 1996; 94(5):992-1002. DOI: 10.1161/01.cir.94.5.992. View

17.

MULIERI L, Hasenfuss G, Leavitt B, Allen P, ALPERT N . Altered myocardial force-frequency relation in human heart failure. Circulation. 1992; 85(5):1743-50. DOI: 10.1161/01.cir.85.5.1743. View

18.

Hinds S, Bian W, Dennis R, Bursac N . The role of extracellular matrix composition in structure and function of bioengineered skeletal muscle. Biomaterials. 2011; 32(14):3575-83. PMC: 3057410. DOI: 10.1016/j.biomaterials.2011.01.062. View

19.

Mummery C, Zhang J, Ng E, Elliott D, Elefanty A, Kamp T . Differentiation of human embryonic stem cells and induced pluripotent stem cells to cardiomyocytes: a methods overview. Circ Res. 2012; 111(3):344-58. PMC: 3578601. DOI: 10.1161/CIRCRESAHA.110.227512. View

20.

Lompre A, Nadal-Ginard B, Mahdavi V . Expression of the cardiac ventricular alpha- and beta-myosin heavy chain genes is developmentally and hormonally regulated. J Biol Chem. 1984; 259(10):6437-46. View