NKX2-5eGFP/w hESCs for isolation of human cardiac progenitors and cardiomyocytes

Elliott, David A.; Braam, Stefan; Koutsis, Katerina; Ng, Elizabeth; Jenny, Robert Alexander; Lagerqvist, Ebba L.; Biben, Christine; Hatzistavrou, Tanya; Hirst, Claire Elizabeth; Yu, Qing; Skelton, Rhys J.P.; Oostwaard, Dorien Ward‐van; Lim, Sue Mei; Khammy, Ouda; Li, Xueling; Hawes, Susan Melanie; Davis, Richard P.; Goulburn, Adam L; Passier, Robert; Prall, Owen W.J.; Haynes, John M.; Pouton, Colin W.; Kaye, David M.; Mummery, Christine L.; Elefanty, Andrew G.

doi:10.1038/nmeth.1740

Cited by 378 publications

(376 citation statements)

References 28 publications

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“…To test the hypothesis that cell composition would affect tissue physiology, and to extend our analysis to the formulation of more translationally relevant human cardiac microtissues, we sorted hPSC-derived heart cells to generate tissues with specific input populations consisting of NKX2-5+ cardiomyocytes and CD90+ nonmyocytes (putatively fibroblasts; FBs). We applied our cardiac differentiation protocol to an NKX2-5-GFP reporter human embryonic stem cell (hESC) line that contained the EGFP cDNA inserted into the NKX2-5-GFP locus of HES3 hESC (1,9). On day 20, we dissociated the hESC-CM aggregates and sorted for both of these populations to high purity (Fig.…”

Section: Input-population Composition Impacts Tissue Morphogenesis Andmentioning

confidence: 99%

“…microfabrication | heart regeneration | tissue engineering | cardiac toxicity | arrhythmia disease model D irected differentiation strategies for generating and preserving human pluripotent stem cell (hPSC)-derived cardiomyocytes (CMs) are well-developed (1)(2)(3)(4)(5)(6)(7), and several CM cell-surface markers that can be used to enrich target subpopulations have been discovered (8,9). Although differentiation of hPSCs into contracting CMs is well-established, their maturation into adult-equivalent cells, and their formulation into functional adult-like tissue, remains an unmet challenge (5).…”

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confidence: 99%

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Design and formulation of functional pluripotent stem cell-derived cardiac microtissues

Thavandiran

Dubois

Mikryukov

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

251

248

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Significance Robust and predictive in vitro models of human cardiac tissue function could have transformative impact on our ability to test new drugs and understand cardiac disease. Despite significant effort, the generation of high-fidelity adult-like human cardiac tissue analogs remains challenging. In this paper, we systematically explore the design criteria for pluripotent stem cell-derived engineered cardiac tissue. Parameters such as biomechanical stress during tissue remodeling, input-cell composition, electrical stimulation, and tissue geometry are evaluated. Our results suggest that a specified combination of a 3D matrix-based microenvironment, uniaxial mechanical stress, and mixtures of cardiomyocytes and fibroblasts improves the performance and maturation state of in vitro engineered cardiac tissue.

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Section: Input-population Composition Impacts Tissue Morphogenesis Andmentioning

confidence: 99%

mentioning

confidence: 99%

Design and formulation of functional pluripotent stem cell-derived cardiac microtissues

Thavandiran

Dubois

Mikryukov

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

251

248

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“…Current procedures for eliminating such contamination and boosting cardiomyocyte enrichment involve genetic modification [14,15] and nongenetic methods using a mitochondrial dye [16] or antibodies to specific cell-surface markers [17]. However, none of these methods are ideal for the therapeutic application of PSC-derived cardiomyocytes due to insufficient stability, genotoxicity, and the use of fluorescence-activated cell sorting (FACS).…”

Section: Nongenetic Methods For Purifying Cardiomyocytesmentioning

confidence: 99%

Future Treatment of Heart Failure Using Human iPSC-Derived Cardiomyocytes

Tohyama

Fukuda

2016

Etiology and Morphogenesis of Congenital Heart Disease

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Heart transplantation can drastically improve survival in patients with a failing heart; however, the shortage of donor hearts remains a serious problem with this treatment strategy and the successful clinical application of regenerative medicine is eagerly awaited. To this end, we developed a novel method to generate human induced pluripotent stem cells (iPSCs) from circulating human T lymphocytes using Sendai virus containing Yamanaka factors. To establish an efficient cardiac differentiation protocol, we then screened factors expressed in the future heart site of early mouse embryos and identified several growth factors and cytokines that can induce cardiomyocyte differentiation and proliferation. Subsequent transcriptome and metabolome analysis on undifferentiated stem cells and cardiomyocytes to devise a specific metabolic environment for cardiomyocyte selection revealed completely different mechanisms of glucose and lactate metabolism. Based on these findings, we succeeded in metabolically selecting cardiomyocytes using glucose-free and lactate-supplemented medium, with up to 99 % purity and no teratoma formation. Using our aggregation technique, we also showed that >90 % of the transplanted cardiomyocytes survived in the heart and showed physiological growth after transplantation. We expect that combining these techniques will achieve future heart regeneration.

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“…animal serums and Matrigel) must not come into contact with cells and cell products that [93]. Several types of specific nutrient mediums devoid of products of undefined and/or animal origin were designed specifically for differentiation of stem cells into the cardiomyocyte lineage [94,95].…”

Section: Cardiomyocytes Derived By Reprogramming Of Somatic Cellsmentioning

confidence: 99%

“…Embryoid body-based methods for derivation of cardiomyocytes use specialised mediums such as APEL (albumin, polyvinyl alcohol, essential lipids) and StemPro-34 [87,95]. Growth factors need to be added to both mediums, allowing for more precise assessment of their effects.…”

Section: Cardiomyocytes Derived By Reprogramming Of Somatic Cellsmentioning

confidence: 99%

We heart cultured hearts. A comparative review of methodologies for targeted differentiation and maintenance of cardiomyocytes derived from pluripotent and multipotent stem cells

Arabadjiev¹,

Petkova²,

Chakarov³

et al. 2014

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Citation: Arabadjiev A, Petkova R, Chakarov S, Pankov R, Zhelev N. We heart cultured hearts. A comparative review of methodologies for targeted differentiation and maintenance of cardiomyocytes derived from pluripotent and multipotent stem cells. AbstractHuman cell lines, including disease cell lines are often superior to routine animal models for the purposes of rapid and safe assessment of the effects of different agents that may modulate myocardial functioning under physiological and pathological conditions. There are several currently existing methodologies for derivation of cardiomyocyte-like cells by targeted differentiation from pluripotent cells and by reprogramming/ transdifferentiation from other types of cells (multipotent progenitors, somatic cells, etc). The present paper reviews the potential sources of cells capable of differentiation along the cardiomyocyte lineage; the existing methodologies for targeted differentiation, outlining the specificities of each method; and the markers for differentiation along the mesodermal and the cardiogenic lineage. The yield of robustly beating cells expressing cardio-specific proteins derived by any of the existing methods, however, still rarely exceeds 70-90 %, even with the newly developed approaches for increasing the differentiation capacity. There still is significant variance in the results obtained by different research groups and even between different experiments carried out in the same laboratory, with the same type of cells and same general type of protocol. Derivation of new lines of human pluripotent and multipotent stem cells according to standardised protocols and in completely defined; xeno-free conditions may increase the reliability and reproducibility of research and speed up the development of potential clinical applications.

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NKX2-5eGFP/w hESCs for isolation of human cardiac progenitors and cardiomyocytes

Cited by 378 publications

References 28 publications

Design and formulation of functional pluripotent stem cell-derived cardiac microtissues

Design and formulation of functional pluripotent stem cell-derived cardiac microtissues

Future Treatment of Heart Failure Using Human iPSC-Derived Cardiomyocytes

We heart cultured hearts. A comparative review of methodologies for targeted differentiation and maintenance of cardiomyocytes derived from pluripotent and multipotent stem cells

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