Isolation and characterization of ventricular-like cells derived from NKX2-5 and MLC2v double knock-in human pluripotent stem cells

Yamauchi, Kaori; Li, Junjun; Morikawa, Kumi; Liu, Li; Shirayoshi, Yasuaki; Nakatsuji, Norio; Elliott, David A.; Hisatome, Ichiro; Suemori, Hirofumi

doi:10.1016/j.bbrc.2017.11.133

Cited by 9 publications

(6 citation statements)

References 22 publications

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“…To be able to isolate ventricular and atrial cardiomyocytes from mixed differentiation cultures, several groups have engineered hPSC lines to express fluorescent reporters or drug selection genes from the MLC2V locus (Huber et al, 2007;Li et al, 2017;Yamauchi et al, 2018) or the COUP-TFII locus, respectively (Schwach et al, 2017). While these approaches allow for the isolation of highly enriched populations, they do have a number of drawbacks that may limit their usefulness for clinical applications.…”

Section: Ventricular and Atrial Cardiomyocytesmentioning

confidence: 99%

Human Pluripotent Stem Cell-Derived Cardiovascular Cells: From Developmental Biology to Therapeutic Applications

Lee²,

2019

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Advances in our understanding of cardiovascular development have provided a roadmap for the directed differentiation of human pluripotent stem cells (hPSCs) to the major cell types found in the heart. In this Perspective, we review the state of the field in generating and maturing cardiovascular cells from hPSCs based on our fundamental understanding of heart development. We then highlight their applications for studying human heart development, modeling disease-performing drug screening, and cell replacement therapy. With the advancements highlighted here, the promise that hPSCs will deliver new treatments for degenerative and debilitating diseases may soon be fulfilled.

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Section: Ventricular and Atrial Cardiomyocytesmentioning

confidence: 99%

Human Pluripotent Stem Cell-Derived Cardiovascular Cells: From Developmental Biology to Therapeutic Applications

Lee²,

2019

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“…For instance, CRISPR/Cas9 editing of hiPSCs has enabled the generation of double reporter TBX5 (T‐Box Transcription Factor 5) Clover2 and NKX2‐5 (NK2 Homeobox 5) TagRFP cells that has been used for the purification of heart cell subtypes, such as cells from the first heart field, epicardial, second heart field, and endothelial lineages [ 80 ]. Alternatively, NKX2‐5 eGFP/w and MLC (Myosin regulatory Light Chain)2v mCherry/w reporter cells have been generated to isolate ventricular like cells [ 81 ]. Genome editing (TALEN) have also been used to introduce a selection marker (neomycin) or GFP after the locus of myosin light chain 2 (MYL2) that can serve for identifying and selecting ventricular cardiomyocytes [ 82 ].…”

Section: Surface Marker Identification For the Isolation Of Hpsc‐deri...mentioning

confidence: 99%

Downstream bioprocessing of human pluripotent stem cell‐derived therapeutics

Sart

Liu

Zeng

et al. 2021

Engineering in Life Sciences

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With the advancement in lineage-specific differentiation from human pluripotent stem cells (hPSCs), downstream cell separation has now become a critical step to produce hPSC-derived products. Since differentiation procedures usually result in a heterogeneous cell population, cell separation needs to be performed either to enrich the desired cell population or remove the undesired cell population. This article summarizes recent advances in separation processes for hPSCderived cells, including the standard separation technologies, such as magneticactivated cell sorting, as well as the novel separation strategies, such as those based on adhesion strength and metabolic flux. Specifically, the downstream bioprocessing flow and the identification of surface markers for various cell lineages are discussed. While challenges remain for large-scale downstream bioprocessing of hPSC-derived cells, the rational quality-by-design approach should be implemented to enhance the understanding of the relationship between process and the product and to ensure the safety of the produced cells.

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“…On the other hand, a stable transgenic hPSC line harboring fluorescent reporter under the transcriptional control of human myosin light chain-2V promoter (MLC2V) [130][131][132] and chick ovalbumin upstream promoter transcription factor II (COUP-TFII) [133] were developed to isolate ventricular and atrial cardiomyocytes, respectively. Despite the robust and high efficiency in enriching specific subtypes after cardiac differentiation of PSCs, the use of virus-based vector, again, has raised safety issues including immunogenicity and insertional mutagenesis risk that hinder their application in future clinical treatment.…”

Section: Diverse Cardiomyocyte Subtypes (Atrial Ventricular and Pacmentioning

confidence: 99%

Mending a broken heart: current strategies and limitations of cell-based therapy

Liew

Soh

2020

Stem Cell Res Ther

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The versatility of pluripotent stem cells, attributable to their unlimited self-renewal capacity and plasticity, has sparked a considerable interest for potential application in regenerative medicine. Over the past decade, the concept of replenishing the lost cardiomyocytes, the crux of the matter in ischemic heart disease, with pluripotent stem cell-derived cardiomyocytes (PSC-CM) has been validated with promising pre-clinical results. Nevertheless, clinical translation was hemmed in by limitations such as immature cardiac properties, long-term engraftment, graft-associated arrhythmias, immunogenicity, and risk of tumorigenicity. The continuous progress of stem cell-based cardiac therapy, incorporated with tissue engineering strategies and delivery of cardio-protective exosomes, provides an optimistic outlook on the development of curative treatment for heart failure. This review provides an overview and current status of stem cell-based therapy for heart regeneration, with particular focus on the use of PSC-CM. In addition, we also highlight the associated challenges in clinical application and discuss the potential strategies in developing successful cardiac-regenerative therapy.

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Isolation and characterization of ventricular-like cells derived from NKX2-5 and MLC2v double knock-in human pluripotent stem cells

Cited by 9 publications

References 22 publications

Human Pluripotent Stem Cell-Derived Cardiovascular Cells: From Developmental Biology to Therapeutic Applications

Human Pluripotent Stem Cell-Derived Cardiovascular Cells: From Developmental Biology to Therapeutic Applications

Downstream bioprocessing of human pluripotent stem cell‐derived therapeutics

Mending a broken heart: current strategies and limitations of cell-based therapy

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