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Author Spotlight: Real-Time Measurements of Calcium and Contractility Parameters in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes
Published on: May 26, 2023
Ca(2+)-Currents in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes Effects of Two Different Culture
Ahmet U Uzun1, Ingra Mannhardt1, Kaja Breckwoldt1
1Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-EppendorfHamburg, Germany; Partner Site Hamburg/Kiel/Lübeck, German Centre for Cardiovascular Research (DZHK)Hamburg, Germany.
Insights
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) in 3D engineered heart tissue (EHT) show improved maturation. EHT cultures enhance L-type calcium channel (ICa,L) function and catecholamine response, mimicking adult cardiomyocyte properties.
Area of Science:
- Cardiology
- Stem Cell Biology
- Biomedical Engineering
Background:
- Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) are crucial for studying heart physiology and disease.
- Assessing hiPSC-CM maturation is vital for their application in regenerative medicine and drug discovery.
- Maturation is often evaluated by comparing hiPSC-CM electrophysiological properties to adult human cardiomyocytes (CM).
Purpose of the Study:
- To investigate if a 3D engineered heart tissue (EHT) culture format promotes hiPSC-CM maturation.
- To compare L-type calcium current (ICa,L) properties in hiPSC-CM cultured in 2D monolayer (ML) versus 3D EHT.
- To assess the functional response of hiPSC-CM to adrenergic and serotonergic stimulation in different culture formats.
Main Methods:
- Patch-clamp electrophysiology was used to measure calcium currents in hiPSC-CM.
- hiPSC-CM were cultured in conventional 2D monolayer (ML) and 3D engineered heart tissue (EHT) formats.
- hiPSC-CM responses to pharmacological agents (e.g., (-)-Bay K 8644, adrenergic agonists) were analyzed and compared to adult CM data.
Main Results:
- hiPSC-CM in EHT exhibited robust L-type calcium currents (ICa,L) and T-type calcium currents (ICa,T).
- EHT culture enhanced the response to β1- and β2-adrenoceptor stimulation, similar to adult CM, unlike ML cultures.
- While basal ICa,L density was comparable to adult CM, EHT cultures showed more efficient maturation, indicated by catecholamine sensitivity.
Conclusions:
- 3D EHT culture promotes maturation of hiPSC-CM, enhancing their physiological relevance.
- hiPSC-CM in EHT exhibit improved adrenergic signaling, crucial for cardiac function.
- EHT represents a superior culture model for hiPSC-CM, facilitating accurate cardiac research and drug screening.
Abstract:
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) provide a unique opportunity to study human heart physiology and pharmacology and repair injured hearts. The suitability of hiPSC-CM critically depends on how closely they share physiological properties of human adult cardiomyocytes (CM). Here we investigated whether a 3D engineered heart tissue (EHT) culture format favors maturation and addressed the L-type Ca(2+)-current (ICa,L) as a readout. The results were compared with hiPSC-CM cultured in conventional monolayer (ML) and to our previous data from human adult atrial and ventricular CM obtained when identical patch-clamp protocols were used. HiPSC-CM were two- to three-fold smaller than adult CM, independently of culture format [capacitance ML 45 ± 1 pF (n = 289), EHT 45 ± 1 pF (n = 460), atrial CM 87 ± 3 pF (n = 196), ventricular CM 126 ± 8 pF (n = 50)]. Only 88% of ML cells showed ICa, but all EHT. Basal ICa density was 10 ± 1 pA/pF (n = 207) for ML and 12 ± 1 pA/pF (n = 361) for EHT and was larger than in adult CM [7 ± 1 pA/pF (p < 0.05, n = 196) for atrial CM and 6 ± 1 pA/pF (p < 0.05, n = 47) for ventricular CM]. However, ML and EHT showed robust T-type Ca(2+)-currents (ICa,T). While (-)-Bay K 8644, that activates ICa,L directly, increased ICa,Lto the same extent in ML and EHT, β1- and β2-adrenoceptor effects were marginal in ML, but of same size as (-)-Bay K 8644 in EHT. The opposite was true for serotonin receptors. Sensitivity to β1 and β2-adrenoceptor stimulation was the same in EHT as in adult CM (-logEC50: 5.9 and 6.1 for norepinephrine (NE) and epinephrine (Epi), respectively), but very low concentrations of Rp-8-Br-cAMPS were sufficient to suppress effects (-logEC50: 5.3 and 5.3 respectively for NE and Epi). Taken together, hiPSC-CM express ICa,L at the same density as human adult CM, but, in contrast, possess robust ICa,T. Increased effects of catecholamines in EHT suggest more efficient maturation.
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