Human iPS cell-engineered cardiac tissue sheets with cardiomyocytes and vascular cells for cardiac regeneration

Masumoto, Hidetoshi; Ikuno, Takeshi; Takeda, Masafumi; Fukushima, Hiroyuki; Marui, Akira; Katayama, Shota; Shimizu, Tatsuya; Ikeda, Tadashi; Okano, Teruo; Sakata, Ryuzo; Yamashita, Jun

doi:10.1038/srep06716

Cited by 237 publications

(207 citation statements)

References 28 publications

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“…From repairing damaged tissue in vivo to predicting drug efficacy, human pluripotent stem cells (hPSCs) have become a promising solution to many bottlenecks in human biological research [1][2][3][4][5][6] . There have been many efforts to generate cardiomyocytes (CMs) derived from human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) due to the difficulty in obtaining human cardiac tissue by other methods 7 .…”

Section: Introductionmentioning

confidence: 99%

Accurate nanoelectrode recording of human pluripotent stem cell-derived cardiomyocytes for assaying drugs and modeling disease

et al. 2017

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The measurement of the electrophysiology of human pluripotent stem cell-derived cardiomyocytes is critical for their biomedical applications, from disease modeling to drug screening. Yet, a method that enables the high-throughput intracellular electrophysiology measurement of single cardiomyocytes in adherent culture is not available. To address this area, we have fabricated vertical nanopillar electrodes that can record intracellular action potentials from up to 60 single beating cardiomyocytes. Intracellular access is achieved by highly localized electroporation, which allows for low impedance electrical access to the intracellular voltage. Herein, we demonstrate that this method provides the accurate measurement of the shape and duration of intracellular action potentials, validated by patch clamp, and can facilitate cellular drug screening and disease modeling using human pluripotent stem cells. This study validates the use of nanopillar electrodes for myriad further applications of human pluripotent stem cell-derived cardiomyocytes such as cardiomyocyte maturation monitoring and electrophysiology-contractile force correlation. There have been many efforts to generate cardiomyocytes (CMs) derived from human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) due to the difficulty in obtaining human cardiac tissue by other methods 7 . In the last decade, efficient and reliable protocols, such as the embryoid body method and the monolayer differentiation method have been established to generate hPSC-CMs within 2 weeks and with 470% efficiency 8,9 . Critically, hPSC-CMs exhibit key cardiomyocyte properties, expressing the expected ion channels, exhibiting human cardiac-type action potentials, and containing functional sarcomeres.Despite their potential, applications of hPSC-CMs have been hampered by the cells' intrinsic heterogeneity and the quality of functional assays for monitoring their electrophysiology. First, hPSC-CMs are intrinsically heterogeneous, consisting of three subpopulations: atrial-, ventricular-, and nodal-like 10 . Furthermore, electrophysiology measurements have shown that the shapes of action potentials vary significantly between studies and within studies among different cell lines and differentiation methods 11,12 . Therefore, the high heterogeneity among hPSC-CMs requires that a large number of cells be analyzed to generate statistically meaningful conclusions.The gold standard for measuring cardiomyocyte electrophysiology is manual patch clamp; however, this technique is laborious

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Section: Introductionmentioning

confidence: 99%

Accurate nanoelectrode recording of human pluripotent stem cell-derived cardiomyocytes for assaying drugs and modeling disease

et al. 2017

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“…Considering that mesoderm cells, which possess the potential to differentiate into cardiovascular cells, peak at differentiation day 5 in our CM differentiation system from hiPSCs, we added VEGF instead of Dkk1 at this time point, and successfully induced vascular cells and CMs simultaneously(CM: 67-85% / EC: 8-13% / MC: 3-19% of total cells; 201B6 line) [31] .…”

Section: Simultaneous Induction Of Cms and Vascular Cells From Humanmentioning

confidence: 99%

Exploiting human iPS cell-derived cardiovascular cell populations toward cardiac regenerative therapy

Masumoto

Yamashita

2016

Stem Cell Transl Investig

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To achieve induced pluripotent stem cell (iPSC)-based cardiac regenerative therapy, it is indispensable to establish a method to efficiently induce cardiovascular cell populations from human iPSCs (hiPSCs). We have previously reported a novel serum-free high-density monolayer culture method to induce cardiomyocytes (CMs) from hiPSCs using a stage-specific sequential treatment with TGFβ superfamily molecules (Activin A and BMP4) and a canonical Wnt antagonist, Dkk1. We also succeeded in purifying the differentiated CMs by cell sorting, immunologically labelling the vascular cell adhesion molecule-1 (VCAM-1), a CM-specific cell surface marker which was determined by cell surface marker screening. To induce vascular cells along with CMs, we modified the CM induction method using vascular endothelial cell growth factor on mesoderm-staged cells, which led to simultaneous induction of CMs and vascular cells. We reassembled the cardiovascular cell populations to form a cell sheet which showed a potential for cardiac functional recovery in a rodent myocardial infarction model. These cell differentiation toward cardiovascular cell populations from hiPSCs and bioengineered transplantation methods, could potentially promote hiPSC-based cardiac regenerative therapy in the future.

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“…Most currently used protocols use direct differentiation of PSC in the monolayer format [59•, 62••] that does not require the formation of EBs and thereby reduces the procedural steps. They are based on Matrigel [66] or feeder-cell coated vessels. The differentiation efficiency is highly depending on the timing, differentiation factors, and culture medium (review in [71]).…”

Section: Cardiac Differentiation Of Pluripotent Stem Cellsmentioning

confidence: 99%

Engineering Cardiovascular Regeneration

et al. 2015

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The human heart has very limited regenerative capacity, making the loss of heart muscle tissue during myocardial infarction essentially irreversible. Regenerative medicine aims at promoting the formation of new functional heart muscle in an injured heart, either by stimulating myocyte proliferation, formation of myocytes from preexisting stem cells, direct reprogramming of fibroblasts into myocytes, or adding new muscle tissue from exogenous stem cell sources. Recent advances in stem cell research make these goals more realistic. This review provides a short overview about different approaches for cardiovascular regeneration, stem cell sources, and modes of application, focusing on current cardiac tissue engineering techniques and their way towards clinical application.

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Human iPS cell-engineered cardiac tissue sheets with cardiomyocytes and vascular cells for cardiac regeneration

Cited by 237 publications

References 28 publications

Accurate nanoelectrode recording of human pluripotent stem cell-derived cardiomyocytes for assaying drugs and modeling disease

Accurate nanoelectrode recording of human pluripotent stem cell-derived cardiomyocytes for assaying drugs and modeling disease

Exploiting human iPS cell-derived cardiovascular cell populations toward cardiac regenerative therapy

Engineering Cardiovascular Regeneration

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