Small molecule Wnt inhibitors enhance the efficiency of BMP-4-directed cardiac differentiation of human pluripotent stem cells.

TY - JOUR

T1 - Small molecule Wnt inhibitors enhance the efficiency of BMP-4-directed cardiac differentiation of human pluripotent stem cells.

AU - Ren, Yongming

AU - Lee, Min Young

AU - Schliffke, Simon

AU - Paavola, Jere

AU - Amos, Peter J

AU - Ge, Xin

AU - Ye, Mingyu

AU - Zhu, Shenjun

AU - Senyei, Grant

AU - Lum, Lawrence

AU - Ehrlich, Barbara E

AU - Qyang, Yibing

PY - 2011

Y1 - 2011

N2 - Human induced pluripotent stem (iPS) cells potentially provide a unique resource for generating patient-specific cardiomyocytes to study cardiac disease mechanisms and treatments. However, existing approaches to cardiomyocyte production from human iPS cells are inefficient, limiting the application of iPS cells in basic and translational cardiac research. Furthermore, strategies to accurately record changes in iPS cell-derived cardiomyocyte action potential duration (APD) are needed to monitor APD-related cardiac disease and for rapid drug screening. We examined whether modulation of the bone morphogenetic protein 4 (BMP-4) and Wnt/?-catenin signaling pathways could induce efficient cardiac differentiation of human iPS cells. We found that early treatment of human iPS cells with BMP-4 followed by late treatment with small molecule Wnt inhibitors led to a marked increase in production of cardiomyocytes compared to existing differentiation strategies. Using immunocytochemical staining and real-time intracellular calcium imaging, we showed that these induced cardiomyocytes expressed typical sarcomeric markers, exhibited normal rhythmic Ca(2+) transients, and responded to both ?-adrenergic and electric stimulation. Furthermore, human iPS cell-derived cardiomyocytes demonstrated characteristic changes in action potential duration in response to cardioactive drugs procainamide and verapamil using voltage-sensitive dye-based optical recording. Thus, modulation of the BMP-4 and Wnt signaling pathways in human iPS cells leads to highly efficient production of cardiomyocytes with typical electrophysiological function and pharmacologic responsiveness. The use of human iPS cell-derived cardiomyocytes and the application of calcium- and voltage-sensitive dyes for the direct, rapid measurement of iPS cell-derived cardiomyocyte activity promise to offer attractive platforms for studying cardiac disease mechanisms and therapeutics.

AB - Human induced pluripotent stem (iPS) cells potentially provide a unique resource for generating patient-specific cardiomyocytes to study cardiac disease mechanisms and treatments. However, existing approaches to cardiomyocyte production from human iPS cells are inefficient, limiting the application of iPS cells in basic and translational cardiac research. Furthermore, strategies to accurately record changes in iPS cell-derived cardiomyocyte action potential duration (APD) are needed to monitor APD-related cardiac disease and for rapid drug screening. We examined whether modulation of the bone morphogenetic protein 4 (BMP-4) and Wnt/?-catenin signaling pathways could induce efficient cardiac differentiation of human iPS cells. We found that early treatment of human iPS cells with BMP-4 followed by late treatment with small molecule Wnt inhibitors led to a marked increase in production of cardiomyocytes compared to existing differentiation strategies. Using immunocytochemical staining and real-time intracellular calcium imaging, we showed that these induced cardiomyocytes expressed typical sarcomeric markers, exhibited normal rhythmic Ca(2+) transients, and responded to both ?-adrenergic and electric stimulation. Furthermore, human iPS cell-derived cardiomyocytes demonstrated characteristic changes in action potential duration in response to cardioactive drugs procainamide and verapamil using voltage-sensitive dye-based optical recording. Thus, modulation of the BMP-4 and Wnt signaling pathways in human iPS cells leads to highly efficient production of cardiomyocytes with typical electrophysiological function and pharmacologic responsiveness. The use of human iPS cell-derived cardiomyocytes and the application of calcium- and voltage-sensitive dyes for the direct, rapid measurement of iPS cell-derived cardiomyocyte activity promise to offer attractive platforms for studying cardiac disease mechanisms and therapeutics.

KW - Animals

KW - Humans

KW - Cells, Cultured

KW - Mice

KW - Biological Markers/metabolism

KW - Gene Expression Regulation, Developmental/drug effects

KW - Calcium/metabolism

KW - Action Potentials/drug effects

KW - Bone Morphogenetic Protein 4/pharmacology

KW - Cell Differentiation/drug effects

KW - Embryonic Stem Cells/cytology/drug effects/metabolism

KW - Induced Pluripotent Stem Cells/cytology/drug effects/metabolism

KW - Mesoderm/drug effects/metabolism

KW - Myocytes, Cardiac/cytology/drug effects/metabolism

KW - Wnt Proteins/antagonists & inhibitors

KW - beta Catenin/genetics/metabolism

KW - Animals

KW - Humans

KW - Cells, Cultured

KW - Mice

KW - Biological Markers/metabolism

KW - Gene Expression Regulation, Developmental/drug effects

KW - Calcium/metabolism

KW - Action Potentials/drug effects

KW - Bone Morphogenetic Protein 4/pharmacology

KW - Cell Differentiation/drug effects

KW - Embryonic Stem Cells/cytology/drug effects/metabolism

KW - Induced Pluripotent Stem Cells/cytology/drug effects/metabolism

KW - Mesoderm/drug effects/metabolism

KW - Myocytes, Cardiac/cytology/drug effects/metabolism

KW - Wnt Proteins/antagonists & inhibitors

KW - beta Catenin/genetics/metabolism

M3 - SCORING: Journal article

VL - 51

SP - 280

EP - 287

JO - J MOL CELL CARDIOL

JF - J MOL CELL CARDIOL

SN - 0022-2828

IS - 3

M1 - 3

ER -