title: Tissue-engineered cardiac patch for advanced functional maturation of human ESC-derived cardiomyocytes.
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.
authors: Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA.
authors: Donghui
authors: D
authors: Zhang
authors: Ilya Y
authors: IY
authors: Shadrin
authors: Jason
authors: J
authors: Lam
authors: Hai-Qian
authors: HQ
authors: Xian
authors: H Ralph
authors: HR
authors: Snodgrass
authors: Nenad
authors: N
authors: Bursac
mesh_terms: D000200:Action Potentials / Q000187:drug effects / Q000235:genetics; D000818:Animals; D002316:Cardiotonic Agents / Q000494:pharmacology; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D051379:Mice; D009200:Myocardial Contraction / Q000187:drug effects / Q000235:genetics; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010641:Phenotype; D011943:Receptors, Adrenergic, beta / Q000378:metabolism; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D002316:Cardiotonic Agents; D011943:Receptors, Adrenergic, beta
references: Circ Res. 2003 Jul 11;93(1):32-9
references: 12791707
references: Stem Cells. 2006 Feb;24(2):236-45
references: 16322641
references: Stem Cells Dev. 2012 Aug 10;21(12):2111-21
references: 22268955
references: Cardiovasc Res. 2009 Sep 1;83(4):688-97
references: 19477968
references: Circ Res. 2012 Jun 8;110(12):1556-63
references: 22570367
references: PLoS One. 2011;6(10):e26397
references: 22028871
references: Am J Physiol Heart Circ Physiol. 2001 Apr;280(4):H1814-20
references: 11247796
references: Circ Res. 2011 Jun 24;109(1):47-59
references: 21597009
references: Ann N Y Acad Sci. 2006 Oct;1080:207-15
references: 17132785
references: J Electrocardiol. 2007 Nov-Dec;40(6 Suppl):S192-6
references: 17993321
references: Nat Protoc. 2009;4(10):1522-34
references: 19798085
references: Proc Natl Acad Sci U S A. 2009 Sep 29;106(39):16568-73
references: 19805339
references: PLoS Comput Biol. 2011 May;7(5):e1002061
references: 21637795
references: Cardiovasc Res. 2012 Jan 1;93(1):50-9
references: 21972180
references: Cell Stem Cell. 2013 Jan 3;12(1):127-37
references: 23168164
references: Circulation. 1996 Sep 1;94(5):992-1002
references: 8790037
references: Circulation. 1992 May;85(5):1743-50
references: 1572031
references: Biomaterials. 2011 May;32(14):3575-83
references: 21324402
references: Circ Res. 2012 Jul 20;111(3):344-58
references: 22821908
references: J Biol Chem. 1984 May 25;259(10):6437-46
references: 6327679
references: Circulation. 1995 Nov 15;92(10):2904-10
references: 7586258
references: Cell Stem Cell. 2011 Feb 4;8(2):228-40
references: 21295278
references: Biomaterials. 2011 Oct;32(30):7514-23
references: 21783246
references: Stem Cells. 2007 Dec;25(12):3038-44
references: 17872499
references: Eur Heart J. 2013 Apr;34(15):1134-46
references: 23103664
references: J Cardiovasc Transl Res. 2011 Oct;4(5):605-15
references: 21744185
references: Tissue Eng Part A. 2012 May;18(9-10):957-67
references: 22115339
references: Eur Heart J. 2013 Sep;34(33):2618-29
references: 22798560
references: Nat Methods. 2011 Oct 23;8(12):1037-40
references: 22020065
references: Nat Protoc. 2013 Jan;8(1):162-75
references: 23257984
references: Biomaterials. 2011 Dec;32(35):9180-7
references: 21906802
references: Nat Methods. 2010 Jan;7(1):61-6
references: 19946277
references: Mol Pharm. 2011 Oct 3;8(5):1495-504
references: 21879736
references: Cell Rep. 2012 Nov 29;2(5):1448-60
references: 23103164
references: Prog Biophys Mol Biol. 2007 May-Jun;94(1-2):144-68
references: 17482242
references: PLoS One. 2011 Apr 01;6(4):e18037
references: 21483779
references: Nat Biotechnol. 2011 Oct 23;29(11):1011-8
references: 22020386
references: Circ Res. 2007 Feb 2;100(2):263-72
references: 17218605
references: Proc Natl Acad Sci U S A. 2012 Jul 3;109(27):E1848-57
references: 22645348
references: Curr Protoc Stem Cell Biol. 2007 Jul;Chapter 1:Unit 1F.2
references: 18785170
references: Circ Res. 2012 Oct 12;111(9):1125-36
references: 22912385