Cardiomyocyte differentiation: Experience and observations from 2 laboratories

Patten, Victoria; Chabaesele, I; Sishi, Balindiwe J.N.; Vuuren, Derick van

doi:10.24170/14-2-2498

Cited by 6 publications

(6 citation statements)

References 23 publications

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“…Despite the latest attempts to optimize PCM isolation, unstable quality and low cell yield are major hurdles that preclude the use of PCMs in nonacute in vitro experiments. , In this scenario, H9c2 surpasses other existing cardiac cell lines (e.g., HL-1, AC16) , and can be further matured toward a cardiomyocyte-like phenotype when treated with retinoic acid (RA). , However, it was brought to our attention that recent studies using H9c2 cells to test distinct biomaterials for cardiac applications did not induce such a cardiomyocyte differentiation, ,− suggesting plausible difficulties to mature H9c2 cells when grown onto engineered substrates. Additionally, other laboratories reported discrepancies when reproducing H9c2 maturation to cardiomyocytes, , indicating some degree of resistance to differentiation. Nevertheless, H9c2 differentiation is imperative to achieve a better resemblance to adult cardiac tissue and perform investigations with higher translational value.…”

Section: Resultsmentioning

confidence: 99%

“…45,46 However, it was brought to our attention that recent studies using H9c2 cells to test distinct biomaterials for cardiac applications did not induce such a cardiomyocyte differentiation, 40,81−84 suggesting plausible difficulties to mature H9c2 cells when grown onto engineered substrates. Additionally, other laboratories reported discrepancies when reproducing H9c2 maturation to cardiomyocytes, 76,85 indicating some degree of resistance to differentiation. Nevertheless, H9c2 differentiation is imperative to achieve a better resemblance to adult cardiac tissue and perform investigations with higher translational value.…”

Section: ■ Introductionmentioning

confidence: 99%

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Conductive Bacterial Nanocellulose-Polypyrrole Patches Promote Cardiomyocyte Differentiation

Srinivasan

Cler

Zapata‐Arteaga

et al. 2023

ACS Appl. Bio Mater.

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The low endogenous regenerative capacity of the heart, added to the prevalence of cardiovascular diseases, triggered the advent of cardiac tissue engineering in the last decades. The myocardial niche plays a critical role in directing the function and fate of cardiomyocytes; therefore, engineering a biomimetic scaffold holds excellent promise. We produced an electroconductive cardiac patch of bacterial nanocellulose (BC) with polypyrrole nanoparticles (Ppy NPs) to mimic the natural myocardial microenvironment. BC offers a 3D interconnected fiber structure with high flexibility, which is ideal for hosting Ppy nanoparticles. BC-Ppy composites were produced by decorating the network of BC fibers (65 ± 12 nm) with conductive Ppy nanoparticles (83 ± 8 nm). Ppy NPs effectively augment the conductivity, surface roughness, and thickness of BC composites despite reducing scaffolds’ transparency. BC-Ppy composites were flexible (up to 10 mM Ppy), maintained their intricate 3D extracellular matrix-like mesh structure in all Ppy concentrations tested, and displayed electrical conductivities in the range of native cardiac tissue. Furthermore, these materials exhibit tensile strength, surface roughness, and wettability values appropriate for their final use as cardiac patches. In vitro experiments with cardiac fibroblasts and H9c2 cells confirmed the exceptional biocompatibility of BC-Ppy composites. BC-Ppy scaffolds improved cell viability and attachment, promoting a desirable cardiomyoblast morphology. Biochemical analyses revealed that H9c2 cells showed different cardiomyocyte phenotypes and distinct levels of maturity depending on the amount of Ppy in the substrate used. Specifically, the employment of BC-Ppy composites drives partial H9c2 differentiation toward a cardiomyocyte-like phenotype. The scaffolds increase the expression of functional cardiac markers in H9c2 cells, indicative of a higher differentiation efficiency, which is not observed with plain BC. Our results highlight the remarkable potential use of BC-Ppy scaffolds as a cardiac patch in tissue regenerative therapies.

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Section: Resultsmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

Conductive Bacterial Nanocellulose-Polypyrrole Patches Promote Cardiomyocyte Differentiation

Srinivasan

Cler

Zapata‐Arteaga

et al. 2023

ACS Appl. Bio Mater.

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“…Multinucleation of H9c2 cells is considered as a morphological indication of its differentiation to cardiac lineage rather than to a rhabdomyocyte lineage 30 . Previous reports have shown the increased expression of cardiac‐specific proteins such as myosin light chain‐2v and cardiac troponin‐T together with a shift in metabolic preference from glycolytic to oxidative metabolism 31 during differentiation towards cardiomyocytes. In this study, the differentiated cells showed increased expression of troponin‐T, myo‐D, and connexin 43 indicative of cardiomyocyte‐lineage specific proteins (Figure 3D).…”

Section: Discussionmentioning

confidence: 99%

A porcine cholecystic extracellular matrix conductive scaffold for cardiac tissue repair

et al. 2022

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Cardiac tissue engineering using cells, scaffolds or signaling molecules is a promising approach for replacement or repair of damaged myocardium. This study addressed the contemporary need for a conductive biomimetic nanocomposite scaffold for cardiac tissue engineering by examining the use of a gold nanoparticle-incorporated porcine cholecystic extracellular matrix for the same. The scaffold had an electrical

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“…10 6 cells were plated and after reaching 80% confluence, differentiation was induced for 7 days by the reduction of fetal bovine serum to 1% and the addition of 1 μmol L −1 of retinoic acid (Sigma-Aldrich) to the culture medium. The differentiation medium was changed every 2 days ( Pereira et al, 2011 ; Suhaeri et al, 2015 ; Patten et al, 2017 ). The cells (cardiomyocytes) were then separated into three groups treated with 100 μmol L −1 H 2 O 2 (Merck) for 1 h ( Oraki Kohshour et al, 2013 ; Ilavenil et al, 2015 ), 10 μmol L −1 of isoproterenol (Sigma-Aldrich) for 48 h ( Chen et al, 2012 ), and 1 μmol L −1 of doxorubicin (Sigma-Aldrich) for 12 h ( Jiang et al, 2020 ).…”

Section: Methodsmentioning

confidence: 99%

Untargeted Metabolomics Studies of H9c2 Cardiac Cells Submitted to Oxidative Stress, β-Adrenergic Stimulation and Doxorubicin Treatment: Investigation of Cardiac Biomarkers

Lima

Amaral²,

Moretto³

et al. 2022

Front. Mol. Biosci.

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One of the biggest challenges in the search for more effective treatments for diseases is understanding their etiology. Cardiovascular diseases (CVD) are an important example of this, given the high number of deaths annually. Oxidative stress (the imbalance between oxidant and antioxidant species in biological system) is one of the factors responsible for CVD occurrence, demanding extensive investigation. Excess of reactive oxygen species (ROS) are primarily responsible for this condition, and clinical and scientific literature have reported a significant increase in ROS when therapeutic drugs, such as doxorubicin and isoproterenol, are administered. In this context, the aim of this study is the investigation of potential biomarkers that might be associated with oxidative stress in cardiomyocytes. For this purpose, H9c2 cardiomyocytes were submitted to oxidative stress conditions by treatment with doxorubicin (DOX), isoproterenol (ISO) and hydrogen peroxide (PER). Metabolomics analyses of the cell extract and the supernatant obtained from the culture medium were then evaluated by CE-ESI(+)-TOF-MS. Following signal processing, statistical analyses, and molecular features annotations, the results indicate changes in the aspartate, serine, pantothenic acid, glycerophosphocholine and glutathione metabolism in the cell extract.

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Cardiomyocyte differentiation: Experience and observations from 2 laboratories

Cited by 6 publications

References 23 publications

Conductive Bacterial Nanocellulose-Polypyrrole Patches Promote Cardiomyocyte Differentiation

Conductive Bacterial Nanocellulose-Polypyrrole Patches Promote Cardiomyocyte Differentiation

A porcine cholecystic extracellular matrix conductive scaffold for cardiac tissue repair

Untargeted Metabolomics Studies of H9c2 Cardiac Cells Submitted to Oxidative Stress, β-Adrenergic Stimulation and Doxorubicin Treatment: Investigation of Cardiac Biomarkers

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