scholarly journals 755 Rat engineered heart tissue is a novel in vitro model to evaluate cardiomyocyte proliferation and fibroblast activation after injury

2021 ◽  
Vol 23 (Supplement_G) ◽  
Author(s):  
Giulio Ciucci ◽  
Karim Rahhali ◽  
Giovanni Cimmino ◽  
Paolo Golino ◽  
Gianfranco Sinagra ◽  
...  
Keyword(s):  
Dna Synthesis ◽  
Proliferative Response ◽  
In Vitro Model ◽  
Neonatal Rat ◽  
Heart Cells ◽  
Collagen Deposition ◽  
Activation Markers ◽  
Custom Made ◽  
Vitro Model

Abstract Aims Adult mammals, including humans, fail to regenerate the majority of the lost cardiomyocytes (CMs) that are replaced with scar tissue after injury. This lack of regenerative response is due to the loss of proliferative capacity of adult CMs which in mice occurs 7 days after birth. An in vitro model that recapitulates these changes has not been developed yet. Using rat engineered heart tissues (rEHTs) we have developed a custom-made cryoinjury system to test the hypothesis that maturation of CMs in EHTs regulates the proliferative response of CMs after injury. Methods rEHT were generated using neonatal rat heart cells. A discrete lesion was produced on the mid-section of mature (Day 18) and immature (Day 6) EHTs using a custom-made system based on liquid nitrogen and a 23G needle and medium was supplemented with EdU for 48 h. Results Cryoinjury in mature EHTs produces a localized injury, preserving their residual contractile activity that does not recover over time. We observed a significant increase of EdU+CMs post injury (6.3 ± 1.9% vs. 10.1 ± 1.6%) without significant changes in Ki67+ and pH3+ CMs suggesting that cryoinjury in mature rEHTs induces DNA synthesis but not CM proliferation. Injury in mature EHTs induced also significant proliferation and activation of fibroblasts with collagen deposition. Interestingly, cryoinjury performed in immature EHTs stimulated a significant proliferative response in CMs Conclusions Similar to adult rodents, we show that cryoinjury induces DNA synthesis in CMs without proliferative response and contractile recovery. On the other hand, cryoinjury in immature EHTs leads to CMs proliferation. Moreover, mature EHT fibroblast response to injury retraces the activation progression of cardiac fibroblast after infarction characterized by proliferation, increase of activation markers, increase of collagen deposition suggesting EHTs as a novel model to investigate the biology of cardiac regeneration upon injury.

scholarly journals Obeticholic acid and INT-767 modulate collagen deposition in a NASH in vitro model

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Beatrice Anfuso ◽  
Claudio Tiribelli ◽  
Luciano Adorini ◽  
Natalia Rosso
Keyword(s):  
In Vitro Model ◽  
Collagen Deposition ◽  
Obeticholic Acid ◽  
Vitro Model

A new in vitro model to detect proarrhythmic effects of drugs on human heart cells

2010 ◽  
Vol 62 (2) ◽  
pp. e17
Author(s):  
Atsushi Sugiyama ◽  
Tetsuo Kitamura ◽  
Fumimasa Nomura ◽  
Tomoyuki Kaneko ◽  
Yuji Nakamura ◽  
...  
Keyword(s):  
Human Heart ◽  
In Vitro Model ◽  
Heart Cells ◽  
Vitro Model

scholarly journals The proarrhythmic features of pathological cardiac hypertrophy in neonatal rat ventricular cardiomyocyte cultures

2020 ◽  
Vol 128 (3) ◽  
pp. 545-553
Author(s):  
Zeinab Neshati ◽  
Martin J. Schalij ◽  
Antoine A. F. de Vries
Keyword(s):  
Action Potential ◽  
Cardiac Hypertrophy ◽  
Action Potential Duration ◽  
In Vitro Model ◽  
Left Ventricular ◽  
Neonatal Rat ◽  
Control Group ◽  
Pathological Cardiac Hypertrophy ◽  
Vitro Model

Different factors may trigger arrhythmias in diseased hearts, including fibrosis, cardiomyocyte hypertrophy, hypoxia, and inflammation. This makes it difficult to establish the relative contribution of each of them to the occurrence of arrhythmias. Accordingly, in this study, we used an in vitro model of pathological cardiac hypertrophy (PCH) to investigate its proarrhythmic features and the underlying mechanisms independent of fibrosis or other PCH-related processes. Neonatal rat ventricular cardiomyocyte (nr-vCMC) monolayers were treated with phorbol 12-myristate 13-acetate (PMA) to create an in vitro model of PCH. The electrophysiological properties of PMA-treated and control monolayers were analyzed by optical mapping at day 9 of culture. PMA treatment led to a significant increase in cell size and total protein content. It also caused a reduction in sarcoplasmic/endoplasmic reticulum Ca2+ ATPase 2 level (32%) and an increase in natriuretic peptide A (42%) and α1-skeletal muscle actin (34%) levels, indicating that the hypertrophic response induced by PMA was, indeed, pathological in nature. PMA-treated monolayers showed increases in action potential duration (APD) and APD dispersion, and a decrease in conduction velocity (CV; APD30 of 306 ± 39 vs. 148 ± 18 ms, APD30 dispersion of 85 ± 19 vs. 22 ± 7 and CV of 10 ± 4 vs. 21 ± 2 cm/s in controls). Upon local 1-Hz stimulation, 53.6% of the PMA-treated cultures showed focal tachyarrhythmias based on triggered activity ( n = 82), while the control group showed 4.3% tachyarrhythmias ( n = 70). PMA-treated nr-vCMC cultures may, thus, represent a well-controllable in vitro model for testing new therapeutic interventions targeting specific aspects of hypertrophy-associated arrhythmias. NEW & NOTEWORTHY Phorbol 12-myristate 13-acetate (PMA) treatment of neonatal rat ventricular cardiomyocytes (nr-vCMCs) led to induction of many significant features of pathological cardiac hypertrophy (PCH), including action potential duration prolongation and dispersion, which provided enough time and depolarizing force for formation of early afterdepolarization (EAD)-induced focal tachyarrhythmias. PMA-treated nr-vCMCs represent a well-controllable in vitro model, which mostly resembles to moderate left ventricular hypertrophy (LVH) rather than severe LVH, in which generation of a reentry is the putative mechanism of its arrhythmias.

The isolated neonatal rat-cardiomyocyte used in an in vitro model for ‘ischemia’. II. Induction of the 68 kDa heat shock protein

1991 ◽  
Vol 1091 (3) ◽  
pp. 278-284 ◽  
Author(s):  
Mark J.M. Tuijl ◽  
Paul M.P. van Bergen en Henegouwen ◽  
Roeland van Wijk ◽  
Arie J. Verkleij
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
In Vitro Model ◽  
Neonatal Rat ◽  
Vitro Model

scholarly journals In Vitro Model to Investigate Communication between Dorsal Root Ganglion and Spinal Cord Glia

2021 ◽  
Vol 22 (18) ◽  
pp. 9725
Author(s):  
Junxuan Ma ◽  
Vaibhav Patil ◽  
Abhay Pandit ◽  
Leo R. Quinlan ◽  
David P. Finn ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Dorsal Root Ganglion ◽  
Conditioned Medium ◽  
Dorsal Root ◽  
In Vitro Model ◽  
Neonatal Rat ◽  
Large Animal ◽  
Large Species ◽  
Vitro Model

Chronic discogenic back pain is associated with increased inflammatory cytokine levels that can influence the proximal peripheral nervous system, namely the dorsal root ganglion (DRG). However, transition to chronic pain is widely thought to involve glial activation in the spinal cord. In this study, an in vitro model was used to evaluate the communication between DRG and spinal cord glia. Primary neonatal rat DRG cells were treated with/without inflammatory cytokines (TNF-α, IL-1β, and IL-6). The conditioned media were collected at two time points (12 and 24 h) and applied to spinal cord mixed glial culture (MGC) for 24 h. Adult bovine DRG and spinal cord cell cultures were also tested, as an alternative large animal model, and results were compared with the neonatal rat findings. Compared with untreated DRG-conditioned medium, the second cytokine-treated DRG-conditioned medium (following medium change, thus containing solely DRG-derived molecules) elevated CD11b expression and calcium signal in neonatal rat microglia and enhanced Iba1 expression in adult bovine microglia. Cytokine treatment induced a DRG-mediated microgliosis. The described in vitro model allows the use of cells from large species and may represent an alternative to animal pain models (3R principles).

scholarly journals Neonatal rat cardiomyocytes as an in vitro model for circadian rhythms in the heart

2017 ◽  
Vol 112 ◽  
pp. 58-63 ◽  
Author(s):  
Bastiaan C. du Pré ◽  
Pieterjan Dierickx ◽  
Sandra Crnko ◽  
Pieter A. Doevendans ◽  
Marc A. Vos ◽  
...  
Keyword(s):  
Circadian Rhythms ◽  
In Vitro Model ◽  
Neonatal Rat ◽  
Rat Cardiomyocytes ◽  
Neonatal Rat Cardiomyocytes ◽  
Vitro Model

Cardiac muscle tissue engineering: toward an in vitro model for electrophysiological studies

1999 ◽  
Vol 277 (2) ◽  
pp. H433-H444 ◽  
Author(s):  
N. Bursac ◽  
M. Papadaki ◽  
R. J. Cohen ◽  
F. J. Schoen ◽  
S. R. Eisenberg ◽  
...  
Keyword(s):  
Cardiac Muscle ◽  
Cardiac Myocytes ◽  
In Vitro Model ◽  
Neonatal Rat ◽  
Electrophysiological Properties ◽  
Impulse Propagation ◽  
Ventricular Cells ◽  
Electrophysiological Studies ◽  
Vitro Model

The objective of this study was to establish a three-dimensional (3-D) in vitro model system of cardiac muscle for electrophysiological studies. Primary neonatal rat ventricular cells containing lower or higher fractions of cardiac myocytes were cultured on polymeric scaffolds in bioreactors to form regular or enriched cardiac muscle constructs, respectively. After 1 wk, all constructs contained a peripheral tissue-like region (50–70 μm thick) in which differentiated cardiac myocytes were organized in multiple layers in a 3-D configuration. Indexes of cell size (protein/DNA) and metabolic activity (tetrazolium conversion/DNA) were similar for constructs and neonatal rat ventricles. Electrophysiological studies conducted using a linear array of extracellular electrodes showed that the peripheral region of constructs exhibited relatively homogeneous electrical properties and sustained macroscopically continuous impulse propagation on a centimeter-size scale. Electrophysiological properties of enriched constructs were superior to those of regular constructs but inferior to those of native ventricles. These results demonstrate that 3-D cardiac muscle constructs can be engineered with cardiac-specific structural and electrophysiological properties and used for in vitro impulse propagation studies.

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