Myocardial commitment from human pluripotent stem cells: Rapid production of human heart grafts

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• 作者：Elena Garreta^a ; ¹ ; Lorena de Oñ ; ate^a ; ^b ; ¹ ; M. Eugenia Ferná ; ndez-Santos^c ; ^d ; Roger Oria^e ; ^f ; Carolina Tarantino^a ; Andreu M. Climent^g ; André ; s Marco^a ; Mireia Samitier^a ; Elena Martí ; nez^h ; Maria Valls-Margarit^h ; Rafael Matesanzⁱ ; Doris A. Taylor^j ; ^k ; ^l ; Francisco Ferná ; ndez-Avilé ; s^c ; ^d ; ^{faviles@secardiologia.es" class="auth_mail" title="E-mail the corresponding author} ; Juan Carlos Izpisua Belmonte^m ; ^{belmonte@salk.edu" class="auth_mail" title="E-mail the corresponding author} ; Nuria Montserrat^a ; ^{nmontserrat@ibecbarcelona.eu" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Gene targeting ; Pluripotent stem cells ; Extracellular matrix ; Cardiac function
• 刊名：Biomaterials
• 出版年：2016
• 出版时间：August 2016
• 年：2016
• 卷：98
• 期：Complete
• 页码：64-78
• 全文大小：6090 K

文摘

Genome editing on human pluripotent stem cells (hPSCs) together with the development of protocols for organ decellularization opens the door to the generation of autologous bioartificial hearts. Here we sought to generate for the first time a fluorescent reporter human embryonic stem cell (hESC) line by means of Transcription activator-like effector nucleases (TALENs) to efficiently produce cardiomyocyte-like cells (CLCs) from hPSCs and repopulate decellularized human heart ventricles for heart engineering. In our hands, targeting myosin heavy chain locus (MYH6) with mCherry fluorescent reporter by TALEN technology in hESCs did not alter major pluripotent-related features, and allowed for the definition of a robust protocol for CLCs production also from human induced pluripotent stem cells (hiPSCs) in 14 days. hPSCs-derived CLCs (hPSCs-CLCs) were next used to recellularize acellular cardiac scaffolds. Electrophysiological responses encountered when hPSCs-CLCs were cultured on ventricular decellularized extracellular matrix (vdECM) correlated with significant increases in the levels of expression of different ion channels determinant for calcium homeostasis and heart contractile function. Overall, the approach described here allows for the rapid generation of human cardiac grafts from hPSCs, in a total of 24 days, providing a suitable platform for cardiac engineering and disease modeling in the human setting.