Generation and characterization of functional cardiomyocytes using induced pluripotent stem cells derived from human fibroblasts.
We have successfully developed both spontaneous and inductive cardiomyocyte differentiation of iPS cells reprogrammed from human foreskin fibroblasts. The reprogrammed iPS cells morphologically resemble human cardiomyocytes which can beat. RT-PCR and immunostaining show that cardiac markers are expressed that are comparable to the differentiation pattern of authentic human embryonic stem cells, indicating the existence of both immature and mature differentiated cardiomyocytes. 5-Azacytidine greatly enhanced the efficiency of cardiomyocyte differentiation, whereas dimethylsulfoxide had no effect. Low serum and bone morphogenetic protein-2 marginally improved differentiation efficiency. iPS cell-derived cardiomyocytes changed their beat frequency in response to cardiac drugs, which included ion channel blockers and alpha/beta adrenergic stimulators. Derived cardiomyocytes look promising as an in vitro system for potential drug screen and/or toxicity, making this system closer to practical use in the near future.
Gai
Hui
H
Division of Laboratory Medicine, Nevada Cancer Institute, Las Vegas, NV 89135, USA.
Leung
Elaine Lai-Han
EL
Costantino
Peter D
PD
Aguila
Jerell R
JR
Nguyen
David M
DM
Fink
Louis M
LM
Ward
David C
DC
Ma
Yupo
Y
D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D001374:Azacitidine / Q000494:pharmacology; D055396:Bone Morphogenetic Protein 2 / Q000378:metabolism / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology; D005347:Fibroblasts / Q000166:cytology* / Q000502:physiology; D052816:Foreskin / Q000166:cytology; D016166:Free Radical Scavengers / Q000494:pharmacology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology; D007473:Ion Channels / Q000187:drug effects / Q000502:physiology; D008297:Male; D009200:Myocardial Contraction; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000502:physiology; D011941:Receptors, Adrenergic / Q000187:drug effects / Q000502:physiology; D014158:Transcription, Genetic
D000964:Antimetabolites, Antineoplastic; D055396:Bone Morphogenetic Protein 2; D016166:Free Radical Scavengers; D007473:Ion Channels; D011941:Receptors, Adrenergic; D001374:Azacitidine; D004121:Dimethyl Sulfoxide