Hyperglycaemia-Induced Contractile Dysfunction and Apoptosis in Cardiomyocyte-Like Pulsatile Cells Derived from Mouse Embryonic Stem Cells

Aboalgasm, Hamida; Ballo, Robea; Mkatazo, Thulisa; Gwanyanya, Asfree

doi:10.1007/s12012-021-09660-3

Cited by 5 publications

(7 citation statements)

References 39 publications

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“…Western blot analysis was performed as previously described (Aboalgasm et al 2021a, Aboalgasm et al 2021b). Sections of frozen LV tissue (approximately 0.05 g) were D r a f t 9 homogenized on ice by sonication in a modified radioimmunoprecipitation assay (RIPA) lysis buffer (50 mM Tris-HCl, 150 mM NaCl, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulphate, pH 7.4) containing a protease/phosphatase inhibitor cocktail (HALT, Thermo Fisher Scientific, Rockford, USA).…”

Section: Western Immunoblottingmentioning

confidence: 99%

Cardioprotective effect of fingolimod against calcium paradox–induced myocardial injury in the isolated rat heart

Alatrag

Amoni

Kelly-Laubscher

et al. 2022

Can. J. Physiol. Pharmacol.

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Fingolimod (FTY720) inhibits Ca2+-permeable, Mg2+-sensitive channels called transient receptor potential melastatin 7 (TRPM7), but its effects on Ca2+ paradox (CP)-induced myocardial damage have not been evaluated. We studied the effect of FTY720 on CP-induced myocardial damage, and used other TRPM7 channel inhibitors nordihydroguaiaretic acid (NDGA) and Mg2+ to test if any effect of FTY720 was via TRPM7 inhibition. Langendorff-perfused Wistar rat hearts were treated with FTY720 or NDGA and subjected to a CP protocol consisting of Ca2+ depletion followed by Ca2+ repletion. Hearts of rats pre-treated with MgSO4 were also subjected to CP. Hemodynamic parameters were measured using an intraventricular balloon, and myocardial infarct size was quantified using triphenyltetrazolium chloride stain. TRPM7 proteins in ventricular tissue were detected using immunoblot analysis. FTY720, but not NDGA, decreased CP-induced infarct size. Both FTY720 and NDGA minimized the CP-induced elevation of left ventricular end-diastolic pressure, but only FTY720 ultimately improved ventricular developed pressure. Mg2+ pre-treatment had effect neither on CP-induced infarct size, hemodynamic parameters during CP, nor the level TRPM7 protein expression in ventricular tissue. Overall, FTY720 attenuated CP-induced myocardial damage, with potential therapeutic implications on Ca2+-mediated cardiotoxicity. However, the cardioprotective mechanism of FTY720 seems to be unrelated to TRPM7 channel modulation.

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Section: Western Immunoblottingmentioning

confidence: 99%

Cardioprotective effect of fingolimod against calcium paradox–induced myocardial injury in the isolated rat heart

Alatrag

Amoni

Kelly-Laubscher

et al. 2022

Can. J. Physiol. Pharmacol.

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“…Cell culture materials were obtained from ThermoFisher Scienti c (LTC Tech, South Africa), unless stated otherwise. Pluripotent mouse embryonic stem cells (mESCs) of the OLA 129 mouse cell line (gifted by Prof. F. Brombacher, University of Cape Town) were used as previously described [16,17]. In short, undifferentiated mESCs were cultured on a mitomycin C inactivated mouse embryonic broblast (iMEF) feeder cell layer on 0.1% gelatin-coated culture dishes under standardised conditions (humidi ed 5% CO 2 at 37°C).…”

Section: Stem Cell Proliferation and Cardiac Differentiationmentioning

confidence: 99%

“…The mESCs were differentiated into cardiomyocyte-like cells using the hanging drop method and embryoid body (EB) formation as previously described [17]. Brie y, to form an EB, a xed number of mESCs (1000 cells; dissociated from the iMEF feeder layer) were seeded into 20 µl of LIF-free differentiation medium [18,19] containing DMEM/25 mM glucose, supplemented with 10% FBS, 1% glutamax, 1% penicillin/streptomycin, and 0.1% βmercaptoethanol.…”

Section: Stem Cell Proliferation and Cardiac Differentiationmentioning

confidence: 99%

“…The mESCs were differentiated in DMEM medium containing either the baseline (25 mM) glucose [20][21][22] or high (50 mM) glucose [23,24] as previously described [16,17], from day one of the differentiation protocol. Osmolality changes due to glucose addition were not adjusted for, as in other in vitro studies [16,17,23,24], and so as to mimic in vivo diabetic hyperglycaemia with uncorrected osmolarity changes. Measurements on EBs on coverslips and EB sample collection were performed after approximately two weeks (day 1719) of the differentiation protocol.…”

Section: Experimental Treatmentsmentioning

confidence: 99%

“…We previously established and characterised a stem cell-derived cardiac cellular model that mimics cardiac cellular development, and showed that hyperglycaemia suppressed the cardiac differentiation of mouse embryonic stem cells (mESCs) and induced cardiomyocyte contractile dysfunction, features that were attributed to oxidative stress and apoptosis [16,17]. However, the effect of hyperglycaemia on the spontaneous beating characteristics of those cardiac-like cells remains unclear.…”

Section: Introductionmentioning

confidence: 99%

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Hyperglycaemia-induced impairment of the autorhythmicity and gap junction activity of mouse embryonic stem cell-derived cardiomyocyte-like cells

Menzele

Aboalgasm

Ballo

et al. 2022

Histochem Cell Biol

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Diabetes mellitus with hyperglycaemia is a major risk factor for malignant cardiac dysrhythmias.However, the underlying mechanisms remain unclear, especially during the embryonic developmental phase of the heart. This study investigated the effect of hyperglycaemia on the pulsatile activity of stem cell-derived cardiomyocytes. Mouse embryonic stem cells (mESCs) were differentiated into cardiac-like cells through embryoid body (EB) formation, in either baseline glucose or high glucose conditions. Action potentials (APs) were recorded using a voltage-sensitive uorescent dye and gap junction activity was evaluated using scrape-loading lucifer yellow dye transfer assay. Molecular components were detected using immunocytochemistry and immunoblot analyses. High glucose decreased the spontaneous beating rate of EBs and shortened the duration of onset of quinidine-induced asystole. Furthermore, it altered AP amplitude, but not AP duration, and had no impact on the expression of the hyperpolarisationactivated cyclic nucleotide-gated isoform 4 (HCN4) channel nor on the EB beating rate response to ivabradine nor isoprenaline. High glucose also decreased both the intercellular spread of lucifer yellow within an EB and the expression of the cardiac gap junction protein connexin 43 as well as upregulated the expression of transforming growth factor beta 1 (TGFβ1) and phosphorylated Smad3. High glucose suppressed the autorhythmicity and gap junction conduction of mESC-derived cardiomyocytes, via mechanisms probably involving TGFβ1/Smad3 signalling. The results allude to glucotoxicity related proarrhythmic effects, with potential clinical implications in foetal diabetic cardiac disease.

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In focus in HCB

Taatjes

Roth

2023

Histochem Cell Biol

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Hyperglycaemia-Induced Contractile Dysfunction and Apoptosis in Cardiomyocyte-Like Pulsatile Cells Derived from Mouse Embryonic Stem Cells

Cited by 5 publications

References 39 publications

Cardioprotective effect of fingolimod against calcium paradox–induced myocardial injury in the isolated rat heart

Cardioprotective effect of fingolimod against calcium paradox–induced myocardial injury in the isolated rat heart

Hyperglycaemia-induced impairment of the autorhythmicity and gap junction activity of mouse embryonic stem cell-derived cardiomyocyte-like cells

In focus in HCB

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