Formation of Cardiac Fibers in Matrigel Matrix

Bakunts, Karina; Gillum, Nikki; Karabekian, Zaruhi; Sarvazyan, Narine

doi:10.2144/000112682

Cited by 29 publications

(25 citation statements)

References 14 publications

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“…matrigel 25,26 ) or artificial substrates (i.e. silicone polymer 27 , organosilane 28 ) positively influences cardiomyocyte adherence, cell viability and cell spreading during the initial plating step and for the subsequent culture period.…”

Section: Pre-platingmentioning

confidence: 99%

Isolation and Culture of Neonatal Mouse Cardiomyocytes

Ehler

Moore-Morris

Lange

2013

JoVE

147

121

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Cultured neonatal cardiomyocytes have long been used to study myofibrillogenesis and myofibrillar functions. Cultured cardiomyocytes allow for easy investigation and manipulation of biochemical pathways, and their effect on the biomechanical properties of spontaneously beating cardiomyocytes.The following 2-day protocol describes the isolation and culture of neonatal mouse cardiomyocytes. We show how to easily dissect hearts from neonates, dissociate the cardiac tissue and enrich cardiomyocytes from the cardiac cell-population. We discuss the usage of different enzyme mixes for cell-dissociation, and their effects on cell-viability. The isolated cardiomyocytes can be subsequently used for a variety of morphological, electrophysiological, biochemical, cell-biological or biomechanical assays. We optimized the protocol for robustness and reproducibility, by using only commercially available solutions and enzyme mixes that show little lot-to-lot variability. We also address common problems associated with the isolation and culture of cardiomyocytes, and offer a variety of options for the optimization of isolation and culture conditions. Video LinkThe video component of this article can be found at

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Section: Pre-platingmentioning

confidence: 99%

Isolation and Culture of Neonatal Mouse Cardiomyocytes

Ehler

Moore-Morris

Lange

2013

JoVE

147

121

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“…Overall, myocytes in these 3D environments maintained spontaneous contractions 2–3 times longer (>45 days) than those on 2D cultures. Neonatal ventricular myocytes have been shown to exhibit regular contractions after being cultured between 3D gel layers of differing stiffnesses and filamentous protein concentrations (Bakunts et al 2008). In addition, more highly organized patterns of myocyte displacement were found in relatively soft fibrinogen gels, bearing bulk shear moduli near those of gels used in current experiments (Shapira-Schweitzer and Seliktar 2007).…”

Section: Discussionmentioning

confidence: 99%

Microdomain heterogeneity in 3D affects the mechanics of neonatal cardiac myocyte contraction

Curtis

Budyn

Desai

et al. 2012

Biomech Model Mechanobiol

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Cardiac muscle cells are known to adapt to their physical surroundings, optimizing intracellular organization and contractile function for a given culture environment. A previously developed in vitro model system has shown that the inclusion of discrete microscale domains (or microrods) in three dimensions (3D) can alter long-term growth responses of neonatal ventricular myocytes. The aim of this work was to understand how cellular contact with such a domain affects various mechanical changes involved in cardiac muscle cell remodeling. Myocytes were maintained in 3D gels over 5 days in the presence or absence of 100 – μm-long microrods, and the effect of this local heterogeneity on cell behavior was analyzed via several imaging techniques. Microrod abutment resulted in approximately twofold increases in the maximum displacement of spontaneously beating myocytes, as based on confocal microscopy scans of the gel xy-plane or the myocyte long axis. In addition, microrods caused significant increases in the proportion of aligned myofibrils (≤20° deviation from long axis) in fixed myocytes. Microrod-related differences in axial contraction could be abrogated by long-term interruption of certain signals of the RhoA-/Rho-associated kinase (ROCK) or protein kinase C (PKC) pathway. Furthermore, microrod-induced increases in myocyte size and protein content were prevented by ROCK inhibition. In all, the data suggest that microdomain heterogeneity in 3D appears to promote the development of axially aligned contractile machinery in muscle cells, an observation that may have relevance to a number of cardiac tissue engineering interventions.

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“…Basic research conducted to explore the viability of cardiac cells inside naturally derived matrices is still ongoing ( Table 2), such is the case of murine cardiac cells cultured within Matrigel, which were capable of behaving in a physiological-like manner, as they formed cardiac-like fibres (Bakunts, Gillum, Karabekian, & Sarvazyan, 2008). Likewise, a collagen hydrogel was shown to support the differentiation and contraction of cardiac myofibroblasts, after being stimulated with transforming growth factor-beta (TGF-β; Lijnen, Petrov, & Fagard, 2003).…”

Section: Cellular Naturally Derived Hydrogels For Cardiovascular Afmentioning

confidence: 99%

Advanced hydrogels for treatment of diabetes

Reyes-Martínez

Ruíz-Pacheco²,

Flores-Valdéz

et al. 2019

J Tissue Eng Regen Med

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Diabetes mellitus is a chronic disease characterized by high levels of glucose in the blood, which leads to metabolic disorders with severe consequences. Today, there is no cure for diabetes. The current management for diabetes and derived medical conditions, such as hyperglycemia, cardiovascular diseases, or diabetic foot ulcer, includes life style changes and hypoglycemia‐based therapy, which do not fully restore euglycemia or the functionality of damaged tissues in patients. This encourages scientists to work outside their boundaries to develop routes that can potentially tackle such metabolic disorders. In this regard, acellular and cellular approaches have represented an alternative for diabetics, although such treatments still face shortcomings related to limited effectiveness and immunogenicity. The advent of biomaterials has brought significant improvements for such approaches, and three‐dimensional extracellular matrix analogs, such as hydrogels, have played a key role in this regard. Advanced hydrogels are being developed to monitor high blood glucose levels and release insulin, as well as serve as a therapeutic technology. Herein, the state of the art in advanced hydrogels for improving treatment of diabetes, from laboratory technology to commercial products approved by drug safety regulatory authorities, will be concisely summarized and discussed.

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Formation of Cardiac Fibers in Matrigel Matrix

Cited by 29 publications

References 14 publications

Isolation and Culture of Neonatal Mouse Cardiomyocytes

Isolation and Culture of Neonatal Mouse Cardiomyocytes

Microdomain heterogeneity in 3D affects the mechanics of neonatal cardiac myocyte contraction

Advanced hydrogels for treatment of diabetes

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