TY - JOUR
T1 - Myocardial commitment from human pluripotent stem cells
T2 - Rapid production of human heart grafts
AU - Garreta, Elena
AU - de Oñate, Lorena
AU - Fernández-Santos, M. Eugenia
AU - Oria, Roger
AU - Tarantino, Carolina
AU - Climent, Andreu M.
AU - Marco, Andrés
AU - Samitier, Mireia
AU - Martínez, Elena
AU - Valls-Margarit, Maria
AU - Matesanz, Rafael
AU - Taylor, Doris A.
AU - Fernández-Avilés, Francisco
AU - Izpisua Belmonte, Juan Carlos
AU - Montserrat, Nuria
N1 - Publisher Copyright:
© 2016 The Author(s).
PY - 2016/8/1
Y1 - 2016/8/1
N2 - 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.
AB - 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.
KW - Cardiac function
KW - Extracellular matrix
KW - Gene targeting
KW - Pluripotent stem cells
UR - http://www.scopus.com/inward/record.url?scp=84968593291&partnerID=8YFLogxK
U2 - 10.1016/j.biomaterials.2016.04.003
DO - 10.1016/j.biomaterials.2016.04.003
M3 - Article
C2 - 27179434
AN - SCOPUS:84968593291
SN - 0142-9612
VL - 98
SP - 64
EP - 78
JO - Biomaterials
JF - Biomaterials
ER -
