Tissue-engineered cardiac patch for advanced functional maturation of human ESC-derived cardiomyocytes

• Published in: Biomaterials Vol. 34; no. 23; pp. 5813 - 5820
• Main Authors: Zhang, Donghui, Shadrin, Ilya Y., Lam, Jason, Xian, Hai-Qian, Snodgrass, H. Ralph, Bursac, Nenad
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ltd 01.07.2013
• Subjects: Action Potentials - drug effects; Action Potentials - genetics; Adrenergic stimulation; Advanced Basic Science; Animals; Cardiac tissue engineering; Cardiotonic Agents - pharmacology; Cell Line; Dentistry; drug therapy; drugs; Embryonic Stem Cells - cytology; Embryonic Stem Cells - drug effects; Embryonic Stem Cells - metabolism; Gene Expression Regulation - drug effects; genes; Human pluripotent stem cells; Humans; Hydrogel; Mice; Myocardial Contraction - drug effects; Myocardial Contraction - genetics; Myocytes, Cardiac - cytology; Myocytes, Cardiac - drug effects; Myocytes, Cardiac - metabolism; Optical mapping; Phenotype; Receptors, Adrenergic, beta - metabolism; sarcomeres; screening; Time Factors; Tissue Engineering - methods; Tissue Scaffolds - chemistry; Adrenergic stimulation; Optical mapping; Hydrogel; Human pluripotent stem cells; Cardiac tissue engineering
• ISSN: 0142-9612; 1878-5905; 1878-5905
• DOI: 10.1016/j.biomaterials.2013.04.026

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/mm2, 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.