Covalent immobilization of luminescent oxygen indicators reduces cytotoxicity

Välimäki, Hannu; Hyvärinen, Tanja; Leivo, Joni; Iftikhar, Haider; Pekkanen-Mattila, Mari; Rajan, Dhanesh Kattipparambil; Verho, Jarmo; Kreutzer, Joose; Ryynänen, Tomi; Pirhonen, Jonatan; Aalto‐Setälä, Katriina; Kallio, Pasi; Narkilahti, Susanna; Lekkala, Jukka

doi:10.1007/s10544-020-00495-3

Cited by 5 publications

(7 citation statements)

References 47 publications

(53 reference statements)

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“…The decrease in cardiomyocyte beating frequency during ischemic stress has been earlier reported in primary CMs from rat 27 – 29 , in perfused whole mouse heart 30 . We and others have also reported the decrease with pluripotent stem cell derived-CMs 11 , 12 , 24 , 31 , as well as the increase in the beating frequency upon reoxygenation 11 , 12 , 24 , 27 , 29 – 31 .…”

Section: Discussionsupporting

confidence: 65%

“…The partial pressure of oxygen (pO 2 ) was measured from one sample to verify that the oxygen dynamics in the cell culture were as expected. In the verification experiment, a luminescence-based sensor together with a highly biocompatible sensing material, developed by Välimäki and coworkers 24 was used to monitor the oxygen level in the culture. The pO 2 was measured once a minute during the entire experiment, while MEA signals were recorded for 1 min every 30 min throughout the experiment at 20 kHz sampling frequency.…”

Section: Methodsmentioning

confidence: 99%

“…Despite their drawbacks, hiPSC-CM based ischemia–reperfusion models have lately emerged 8 – 23 and these models have been reported to recapitulate the typical cardiac ischemia responses in the cells, such as increased cell death 8 – 10 and disruption of the sarcomere structure 9 , 10 . Furthermore, changes in the functionality and electrophysiology have been observed, including decrease in hiPSC-CM beating frequency 11 , 12 , 15 , 16 , 24 , contractility, calcium overload and arrhythmias 14 . In these models, hypoxic conditions have been induced to the hiPSC-CMs by multiple methods, such as by using hypoxic gas 8 – 10 , 13 – 15 , 24 , 25 or causing oxidative stress via peroxide treatment 16 .…”

Section: Introductionmentioning

confidence: 99%

“…Furthermore, changes in the functionality and electrophysiology have been observed, including decrease in hiPSC-CM beating frequency 11 , 12 , 15 , 16 , 24 , contractility, calcium overload and arrhythmias 14 . In these models, hypoxic conditions have been induced to the hiPSC-CMs by multiple methods, such as by using hypoxic gas 8 – 10 , 13 – 15 , 24 , 25 or causing oxidative stress via peroxide treatment 16 . In addition, cell culture media composition has been modified to better recapitulate the ischemic conditions in several studies.…”

Section: Introductionmentioning

confidence: 99%

“…In this study, we present a hiPSC-CM based platform for modeling cardiac ischemia–reperfusion. The base of the platform is an OxyGenie mini-incubator 26 combined with a microelectrode array (MEA) and a luminescence-based oxygen sensor 12 , 24 . The system allows precise control over oxygen concentration, real-time monitoring of the cardiomyocyte functionality as well as the measurement of oxygen level in the cell culture area during hypoxia and reoxygenation without disturbing or interrupting the experiment.…”

Section: Introductionmentioning

confidence: 99%

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Human induced pluripotent stem cell-based platform for modeling cardiac ischemia

Häkli

Kreutzer

Mäki

et al. 2021

Sci Rep

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Ischemic heart disease is a major cause of death worldwide, and the only available therapy to salvage the tissue is reperfusion, which can initially cause further damage. Many therapeutics that have been promising in animal models have failed in human trials. Thus, functional human based cardiac ischemia models are required. In this study, a human induced pluripotent stem cell derived-cardiomyocyte (hiPSC-CM)-based platform for modeling ischemia–reperfusion was developed utilizing a system enabling precise control over oxygen concentration and real-time monitoring of the oxygen dynamics as well as iPS-CM functionality. In addition, morphology and expression of hypoxia-related genes and proteins were evaluated as hiPSC-CM response to 8 or 24 h hypoxia and 24 h reoxygenation. During hypoxia, initial decrease in hiPSC-CM beating frequency was observed, after which the CMs adapted to the conditions and the beating frequency gradually increased already before reoxygenation. During reoxygenation, the beating frequency typically first surpassed the baseline before settling down to the values close the baseline. Furthermore, slowing on the field potential propagation throughout the hiPSC-CM sheet as well as increase in depolarization time and decrease in overall field potential duration were observed during hypoxia. These changes were reversed during reoxygenation. Disorganization of sarcomere structures was observed after hypoxia and reoxygenation, supported by decrease in the expression of sarcomeric proteins. Furthermore, increase in the expression of gene encoding glucose transporter 1 was observed. These findings indicate, that despite their immature phenotype, hiPSC-CMs can be utilized in modeling ischemia–reperfusion injury.

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Section: Discussionsupporting

confidence: 65%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Human induced pluripotent stem cell-based platform for modeling cardiac ischemia

Häkli

Kreutzer

Mäki

et al. 2021

Sci Rep

Self Cite

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Electrophysiological Changes of Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes during Acute Hypoxia and Reoxygenation

Häkli

Kreutzer

Mäki

et al. 2022

Stem Cells International

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Ischemic heart disease is the most common cardiovascular disease and a major burden for healthcare worldwide. However, its pathophysiology is still not fully understood, and human-based models for disease mechanisms and treatments are needed. Here, we used human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to model acute ischemia-reperfusion in our novel cell culture assembly. The assembly enables exchange of oxygen partial pressure for the cells within minutes, mimicking acute ischemic event. In this study, hypoxia was induced using 0% O2 gas for three hours and reoxygenation with 19% O2 gas for 24 hours in serum- and glucose-free medium. According to electrophysiological recordings, hypoxia decreased the hiPSC-CM-beating frequency and field potential (FP) amplitude. Furthermore, FP depolarization time and propagation slowed down. Most of the electrophysiological changes reverted during reoxygenation. However, immunocytochemical staining of the hypoxic and reoxygenation samples showed that morphological changes and changes in the sarcomere structure did not revert during reoxygenation but further deteriorated. qPCR results showed no significant differences in apoptosis or stress-related genes or in the expression of glycolytic genes. In conclusion, the hiPSC-CMs reproduced many characteristic changes of adult CMs during ischemia and reperfusion, indicating their usefulness as a human-based model of acute cardiac ischemia-reperfusion.

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BEaTS-β: an open-source electromechanical bioreactor for simulating human cardiac disease conditions

Takaya,

Comtois-Bona,

Spasojevic

et al. 2023

Front. Bioeng. Biotechnol.

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Heart disease remains the leading cause of worldwide mortality. Although the last decades have broadened our understanding of the biology behind the pathologies of heart disease, ex vivo systems capable of mimicking disease progression and abnormal heart function using human cells remain elusive. In this contribution, an open-access electromechanical system (BEaTS-β) capable of mimicking the environment of cardiac disease is reported. BEaTS-β was designed using computer-aided modeling to combine tunable electrical stimulation and mechanical deformation of cells cultured on a flexible elastomer. To recapitulate the clinical scenario of a heart attack more closely, in designing BEaTS-β we considered a device capable to operate under hypoxic conditions. We tested human induced pluripotent stem cell-derived cardiomyocytes, fibroblasts, and coronary artery endothelial cells in our simulated myocardial infarction environment. Our results indicate that, under simulated myocardium infarction, there was a decrease in maturation of cardiomyocytes, and reduced survival of fibroblasts and coronary artery endothelial cells. The open access nature of BEaTS-β will allow for other investigators to use this platform to investigate cardiac cell biology or drug therapeutic efficacy in vitro under conditions that simulate arrhythmia and/or myocardial infarction.

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Covalent immobilization of luminescent oxygen indicators reduces cytotoxicity

Cited by 5 publications

References 47 publications

Human induced pluripotent stem cell-based platform for modeling cardiac ischemia

Human induced pluripotent stem cell-based platform for modeling cardiac ischemia

Electrophysiological Changes of Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes during Acute Hypoxia and Reoxygenation

BEaTS-β: an open-source electromechanical bioreactor for simulating human cardiac disease conditions

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