Efficient Generation of Cardiac Purkinje Cells from ESCs by Activating cAMP Signaling

Tsai, Su-Yi; Maass, Karen; Jia, Lan; Fishman, Glenn I.; Chen, Shuibing; Evans, Todd

doi:10.1016/j.stemcr.2015.04.015

Cited by 33 publications

(27 citation statements)

References 46 publications

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“…Without positive or negative selection, 50–70% of these hiPSC-CMs are ventricular or ventricular-like cells, with the remaining cells being either atrial or nodal subtypes 35 . Methods for directed differentiation and selection of atrial 36 , ventricular 37 , and even cardiac purkinje 38 , subtypes of iPSC-CMs have been described, but these have not been applied in most reported studies. Therefore cellular heterogeneity must be considered when interpreting data from hiPSC-CM experiments.…”

Section: Electrophysiologic Characteristics Of Hipsc-cms As Compared mentioning

confidence: 99%

Modeling Inherited Arrhythmia Disorders Using Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Bezzerides

Zhang

2017

Circ J

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Inherited arrhythmia disorders are a group of potentially lethal diseases that remain diagnostic and management challenges. While the genetic basis for many of these disorders is well known, the pathogenicity of individual mutations and the resulting clinical outcomes are difficult to predict. Treatment options remain imperfect, and optimizing therapy for individual patients can be difficult. Recent advances in the derivation of induced pluripotent stem cells (iPSCs) from patients and creation of genetically engineered human models using CRISPR/Cas9 has the potential to dramatically advance translational arrhythmia research. In this review, we discuss the current state of modeling inherited arrhythmia disorders using human iPSC-derived cardiomyocytes. We also discuss current limitations and areas for further study.

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Section: Electrophysiologic Characteristics Of Hipsc-cms As Compared mentioning

confidence: 99%

Modeling Inherited Arrhythmia Disorders Using Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Bezzerides

Zhang

2017

Circ J

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“…These include “primary” chemical messengers, especially steroid hormones (e.g., estrogens, androgens, and glucocorticosteroids) and growth factors (e.g., EGF, FGF, TGF‐β, activin‐A, BMP‐4, HGF, βNGF) (Hong, Zhang, Sultana, Yu, & Wei, ; Huang et al, ; Schuldiner, Yanuka, Itskovitz‐Eldor, Melton, & Benvenisty, ). “Secondary” chemical factors include intracellular messengers, such as cyclic nucleotides (Tsai et al, ). In addition, post‐transciptional regulators such as microRNAs (e.g., mi302/367, 9 and 124) are involved (Chen et al, ).…”

Section: Introductionmentioning

confidence: 99%

Mesenchymal stem cell differentiation: Control by calcium‐activated potassium channels

Pchelintseva

Djamgoz

2017

Journal Cellular Physiology

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Mesenchymal stem cells (MSCs) are widely used in modern medicine for which understanding the mechanisms controlling their differentiation is fundamental. Ion channels offer novel insights to this process because of their role in modulating membrane potential and intracellular milieu. Here, we evaluate the contribution of calcium-activated potassium (K ) channels to the three main components of MSC differentiation: initiation, proliferation, and migration. First, we demonstrate the importance of the membrane potential (V ) and the apparent association of hyperpolarization with differentiation. Of K subtypes, most evidence points to activity of big-conductance channels in inducing initiation. On the other hand, intermediate-conductance currents have been shown to promote progression through the cell cycle. While there is no information on the role of K channels in migration of MSCs, work from other stem cells and cancer cells suggest that intermediate-conductance and to a lesser extent big-conductance channels drive migration. In all cases, these effects depend on species, tissue origin and lineage. Finally, we present a conceptual model that demonstrates how K activity could influence differentiation by regulating V and intracellular Ca oscillations. We conclude that K channels have significant involvement in MSC differentiation and could potentially enable novel tissue engineering approaches and therapies.

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“…Statistical significance is determined using a two-tailed Student's t-test (P <0.05). Primer sequences are listed in Tsai et al (2015).…”

Section: Flow Cytometrymentioning

confidence: 99%

“…For instance, we performed immunofluorescence staining to detect CCS markers, electrophysiologybased experiments to analyze cell identity, fluorescence-activated cell sorting (FACS) analysis to determine the CCS differentiation efficiency and sort ESC-derived cardiac PC, and quantitative real-time PCR to examine the expression profiles of derived cardiac PC. The protocols presented below are detailed descriptions extended from a published article (Tsai et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Efficient Generation of Cardiac Purkinje‐like Cells from Embryonic Stem Cells by Activating cAMP Signaling

Tsai

Chen

Evans

2017

CP Stem Cell Biology

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Strategies to derive cardiac conduction system (CCS) cells including Purkinje cells (PC) would facilitate models for mechanistic studies and drug discovery, and also provide new cellular materials for regenerative therapies. However, using current cardiac differentiation protocols, the differentiation efficiency of CCS cells is extremely low, typically below 1% of the culture. High-throughput chemical screening is a powerful strategy for identifying small molecules that can activate signaling pathways to enhance embryonic stem cell (ESC) differentiation. We describe how to carry out a high- throughput screen to identify small molecules that can efficiently promote CCS generation from mouse ESCs. We also describe several assays, including immunofluorescence staining, electrophysiology, FACS analysis and quantitative real- time PCR, to characterize the phenotype of ESC-derived PC. In summary, we describe an efficient way to identify small molecules that enhance cardiac PC generation. These protocols can be adapted to identify other rare cell lineages by directed differentiation from ESCs. © 2017 by John Wiley & Sons, Inc.

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Efficient Generation of Cardiac Purkinje Cells from ESCs by Activating cAMP Signaling

Cited by 33 publications

References 46 publications

Modeling Inherited Arrhythmia Disorders Using Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Modeling Inherited Arrhythmia Disorders Using Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Mesenchymal stem cell differentiation: Control by calcium‐activated potassium channels

Efficient Generation of Cardiac Purkinje‐like Cells from Embryonic Stem Cells by Activating cAMP Signaling

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