Investigating the Role of Contraction during Myofibril Formation using Human Stem Cell-Derived Cardiomyocytes

Yoo, Dasom; Tu, An-Yue; Fenix, Aidan M.; Regnier, Michael

doi:10.1016/j.bpj.2020.11.1644

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“…Undifferentiated human induced pluripotent stem cells (hiPSCs) (WTC11, Male) were maintained on Matrigel (Corning) coated tissue culture plates in mTeSR1 (Stemcell Technologies). Cardiomyocyte-directed differentiation was performed using the RBA-based modified method as previously described 81 . Spontaneous contraction was observed on day 8 post-induction.…”

Section: Methodsmentioning

confidence: 99%

Far-red and sensitive sensor for monitoring real time H₂O₂dynamics with subcellular resolution and in multi-parametric imaging applications

Lee,

Nguyen,

Jin

et al. 2024

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H2O2is a key oxidant in mammalian biology and a pleiotropic signaling molecule at the physiological level, and its excessive accumulation in conjunction with decreased cellular reduction capacity is often found to be a common pathological marker. Here, we present a red fluorescent Genetically Encoded H2O2Indicator (GEHI) allowing versatile optogenetic dissection of redox biology. Our new GEHI, oROS-HT, is a chemigenetic sensor utilizing a HaloTag and Janelia Fluor (JF) rhodamine dye as fluorescent reporters. We developed oROS-HT through a structure-guided approach aided by classic protein structures and recent protein structure prediction tools. Optimized with JF635, oROS-HT is a sensor with 635 nm excitation and 650 nm emission peaks, allowing it to retain its brightness while monitoring intracellular H2O2dynamics. Furthermore, it enables multi-color imaging in combination with blue-green fluorescent sensors for orthogonal analytes and low auto-fluorescence interference in biological tissues. Other advantages of oROS-HT over alternative GEHIs are its fast kinetics, oxygen-independent maturation, low pH sensitivity, lack of photo-artifact, and lack of intracellular aggregation. Here, we demonstrated efficient subcellular targeting and how oROS-HT can map inter and intracellular H2O2diffusion at subcellular resolution. Lastly, we used oROS-HT with the green fluorescent calcium indicator Fluo-4 to investigate the transient effect of the anti-inflammatory agent auranofin on cellular redox physiology and calcium levels via multi-parametric, dual-color imaging.

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Section: Methodsmentioning

confidence: 99%

Far-red and sensitive sensor for monitoring real time H₂O₂dynamics with subcellular resolution and in multi-parametric imaging applications

Lee,

Nguyen,

Jin

et al. 2024

Preprint

Self Cite

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Investigating the Role of Contraction during Myofibril Formation using Human Stem Cell-Derived Cardiomyocytes

Abstract: All the functions of microtubules are driven by dynamic instability, the dominant feature of its assembly dynamics. Catastrophe, the switching 254a Thursday,

Cited by 1 publication

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Far-red and sensitive sensor for monitoring real time H₂O₂dynamics with subcellular resolution and in multi-parametric imaging applications

Far-red and sensitive sensor for monitoring real time H₂O₂dynamics with subcellular resolution and in multi-parametric imaging applications

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Investigating the Role of Contraction during Myofibril Formation using Human Stem Cell-Derived Cardiomyocytes

Abstract: All the functions of microtubules are driven by dynamic instability, the dominant feature of its assembly dynamics. Catastrophe, the switching 254a Thursday,

Cited by 1 publication

References 0 publications

Far-red and sensitive sensor for monitoring real time H2O2dynamics with subcellular resolution and in multi-parametric imaging applications

Far-red and sensitive sensor for monitoring real time H2O2dynamics with subcellular resolution and in multi-parametric imaging applications

Contact Info

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Far-red and sensitive sensor for monitoring real time H₂O₂dynamics with subcellular resolution and in multi-parametric imaging applications

Far-red and sensitive sensor for monitoring real time H₂O₂dynamics with subcellular resolution and in multi-parametric imaging applications