Plasticity of Calcium Signaling Cascades in Human Embryonic Stem Cell-Derived Neural Precursors

Forostyak, Oksana; Romanyuk, Nataliya; Verkhratsky, Alexei; Syková, Eva; Dayanithi, Govindan

doi:10.1089/scd.2012.0624

Cited by 28 publications

(43 citation statements)

References 79 publications

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“…19 We then measured the intracellular calcium responses to ATP and carbachol in differentiated neurons plated in commercial PDL-coated 384-well plates with the rhVTN-N-supplemented medium. The EC 50 of the ATP and carbachol were 9.8 and 55.7nM, respectively (Fig.…”

Section: Resultsmentioning

confidence: 99%

One-Step Seeding of Neural Stem Cells with Vitronectin-Supplemented Medium for High-Throughput Screening Assays

Dai

Long

et al. 2016

SLAS Discovery

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Human neuronal cells differentiated from induced pluripotent cells have emerged as a new model system for the study of disease pathophysiology and evaluation of drug efficacy. Differentiated neuronal cells are more similar in genetics and biological content to the human brain cells than other animal disease models. However, culture of neuronal cells in assay plates requires a labor-intensive procedure of plate pre-coating, hampering its applications in high throughput screening (HTS). We developed a simplified method with one-step seeding of neural stem cells in assay plates by supplementing the medium with a recombinant human vitronectin (VTN), thus avoiding plate pre-coating. Robust results were obtained from cell viability, calcium response, and neurite outgrowth assays using this new method. Our data demonstrate that this approach greatly simplifies high throughput assays using neuronal cells differentiated from human stem cells for translational research.

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Section: Resultsmentioning

confidence: 99%

One-Step Seeding of Neural Stem Cells with Vitronectin-Supplemented Medium for High-Throughput Screening Assays

Dai

Long

et al. 2016

SLAS Discovery

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“…While cells and tissues differentiated from PSCs have been widely used for calcium signaling studies, we found only a limited number of data about such signaling in pluripotent stem cells, and even these data are mainly from mouse cell studies. Calcium signals in hESC-derived cardiomyocytes and neural cell types have been measured in most cases by calcium sensitive dyes: for cardiomyocytes Fura-2/AM [25][26][27][28], Fluo-4/AM [29,30] or dsRed+ [27], while for neural cell types Fluo-3/AM [8,31] Fura-2/AM [8,32] or Oregon green 488 Bapta-1 AM [33] have been applied. In pluripotent cells the application of Fluo-3/AM (mESC- [34], hESC- [8]) or Fluo-4/AM (mESC- [35][36][37], and hESC [12]) has been reported.…”

Section: Methods For Studying Calcium Signals In Human Ps Cellsmentioning

confidence: 99%

“…Generation of suitable human NPCs for neural differentiation and further transplantation is challenging because of the lack of standardized protocols and functional studies. Forostyak et al [32] and Viero et al [69] demonstrated that calcium signals in hPSC derived NPCs may reflect their regeneration capacity after transplantation. These NPCs exhibited spontaneous calcium oscillations and calcium signals evoked by high K+, ATP, glutamate, g-aminobutyric acid (GABA) and caffeine, indicating that the electrophysiological and calcium handling properties of NPCs are similar to those of matured neurons.…”

Section: Q7mentioning

confidence: 99%

Calcium signaling in human pluripotent stem cells

2016

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a b s t r a c tHuman pluripotent stem cells provide new tools for developmental and pharmacological studies as well as for regenerative medicine applications. Calcium homeostasis and ligand-dependent calcium signaling are key components of major cellular responses, including cell proliferation, differentiation or apoptosis. Interestingly, these phenomena have not been characterized in detail as yet in pluripotent human cell sates. Here we review the methods applicable for studying both short-and long-term calcium responses, focusing on the expression of fluorescent calcium indicator proteins and imaging methods as applied in pluripotent human stem cells. We discuss the potential regulatory pathways involving calcium responses in hPS cells and compare these to the implicated pathways in mouse PS cells. A recent development in the stem cell field is the recognition of so called "naïve" states, resembling the earliest potential forms of stem cells during development, as well as the "fuzzy" stem cells, which may be alternative forms of pluripotent cell types, therefore we also discuss the potential role of calcium homeostasis in these PS cell types.

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“…To confront muscle cell diversity by considering how the activation mechanisms for generating Ca 2+ signals have been adapted to control the different cell functions, we utilized the intracellular free Ca 2+ measurement on single cells. For that purpose, a fluorescence microspectrofluorimetry was adapted so that the control and test solutions could be applied using a temperature controlled multichannel polypropylene capillary perfusion system (Forostyak et al 2013(Forostyak et al , 2016a(Forostyak et al , 2016b. A single outlet capillary tubing (100 μm inner diameter) with a flow rate of 250 μl/min was positioned close to the tested cell (<0.5 mm) and thus, the selected cell could be subjected to a constant flow of control buffer or test solutions.…”

Section: Introductionmentioning

confidence: 99%

Analysis of Ca(2+)signaling mechanisms – our experience on the intercellular communication in muscle remodeling

Filip

Mokrý²,

Forostyak³

et al. 2019

Physiol Res

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The aim of this study was to evaluate cell diversity by considering how Ca(2+) signaling has been adapted in skeletal muscle cell function. We characterized single C2C12 myoblasts through intracellular Ca(2+) signaling kinetics after exposure to specific drugs and calcium blockers using fast fluorescence microspectrofluorimetry followed by ATP effect analysis, which confirmed the expression of functional purinergic adenosine and P2 receptors. Further, we found that glutamate sensitivity of C2C12 cells was mediated by ionotropic glutamate receptors; on the other hand, most cells were responsive to cyclopiazonic acid, which inhibits the sarco-endoplasmic reticulum Ca(2+)-ATPase pump. These results suggest that C2C12 cells possess functional L- and P/Q-type voltage-operated Ca2+ channels, ryanodine receptors and functional sarcoplasmic reticulum Ca2+ stores (typical for muscle cells), adenosine and P2 purinergic receptors, as well as ionotropic glutamate receptors. The evaluation of intracellular Ca2+ signaling is a promising approach towards a better understanding and control of the physiopathological properties of myogenic cells that could be used as a predictive factor in the selection of optimal cells for scaffold recellularization or for tissue engineered constructs used in stem cell therapy.

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Plasticity of Calcium Signaling Cascades in Human Embryonic Stem Cell-Derived Neural Precursors

Cited by 28 publications

References 79 publications

One-Step Seeding of Neural Stem Cells with Vitronectin-Supplemented Medium for High-Throughput Screening Assays

One-Step Seeding of Neural Stem Cells with Vitronectin-Supplemented Medium for High-Throughput Screening Assays

Calcium signaling in human pluripotent stem cells

Analysis of Ca(2+)signaling mechanisms – our experience on the intercellular communication in muscle remodeling

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