Contractile activity and cell-cell contact regulate myofibrillar organization in cultured cardiac myocytes.

Simpson, David G.; Decker, Marlene L.; Clark, William; Decker, Robert S.

doi:10.1083/jcb.123.2.323

Cited by 80 publications

(46 citation statements)

References 47 publications

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“…They demonstrated, by cytofluorimetric analysis, that CDCs c-kit+ cells were present in culture with CD90+ and CD31+ subpopulations [15]. We noticed that some cells did not acquire the differentiated phenotype, which supports the idea that there might be different cellular types that provide physical cell-to-cell contact ( Figure 3 already hypothesized by other groups [31,35]. Our results show that the c-kit positive cells differentiated initially into smooth muscle cells and subsequently into skeletal or cardiac myocytes.…”

Section: Cells' Differentiationsupporting

confidence: 70%

C-Kit Positive Cells from Failing Human Hearts: Role of Culturing Media on Cardiomyogenic Potentials

Paparella¹

2015

J Cardiovasc Med Cardiol

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Background: The possibility of culturing heart cells in order to regenerate damaged tissue is a challenging problem. Recent observations have demonstrated the possibility of isolating and expanding resident cardiac stem cells, which could favor regeneration and functional improvement of the myocardial tissue. Aims:To investigate two different culturing media: one promoting c-kit cells' growth and the other promoting differentiation in cardiac muscle cells. Methods:We obtained primary cultures from left ventricle myocardial tissues of 10 human hearts of patients with end-stage heart failure who received heart transplantation. Cells were first cultured in a medium containing high serum and low calcium/magnesium (Ca 2+ /Mg 2+ ) to promote cell growth (medium A). Than they were cultured in another medium that contained lower serum concentration and a variety of different factors in order to induce cell differentiation (medium B). The presence of c-kit, specific for stem cells, α-sarcomeric actin (SA), specific for skeletal and cardiac muscle cells, and α-smooth muscle actin (SMA), specific for smooth muscle cells was studied by immune-cytochemical analysis.Results: A high percentage of c-kit+, SMA-, SA -cells was observed in medium A; in medium B with lower serum and higher Ca 2+ /Mg 2+ concentrations cells became c-kit-, AML+, SA+.In medium A, 78% of the cells were positive for c-kit. After culturing the same cell populations in medium B with lower serum and higher Ca 2+ /Mg 2+ concentrations, the percentage of c-kit positive cells decreased to 21% while the cells positive for SMA and for SA increased respectively from 28 to 82% and from 0 to 59%. Conclusions:Our results confirm the presence of a high percentage of c-kit positive cells in failing human myocardium and, for the first time, suggest a key role of calcium/magnesium concentration in promoting both c-kit cells' growth and their differentiation in human cardiac muscle.

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Section: Cells' Differentiationsupporting

confidence: 70%

C-Kit Positive Cells from Failing Human Hearts: Role of Culturing Media on Cardiomyogenic Potentials

Paparella¹

2015

J Cardiovasc Med Cardiol

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“…Focal adhesion assembly is in part mediated by the mechanical forces placed on the cell, as both stretch and contractility increase the number and size of cardiomyocyte focal adhesions (112,114). The modulation of focal adhesion assembly/disassembly in response to mechanical load may be related to a primary role for focal adhesion assembly in myofibrillogenesis.…”

Section: Focal Adhesion Complexes In Cultured Cardiomyocytesmentioning

confidence: 99%

“…In cultured cardiomyocytes that retain contractile activity, or are stimulated to contract in culture, focal adhesion proteins also reassemble along the cell-substratum interface in register with overlying Z discs of the remodeled sarcomeres. These basal, costameric attachment sites, as well as the remodeled adherens junctions derived from the remaining components of the intercalated disc (114), provide strong cell adhesion sites that retain mechanical communication between the ECM and the contractile machinery of the cultured cell.…”

Section: Focal Adhesion Complexes In Cultured Cardiomyocytesmentioning

confidence: 99%

“…may be considered a striated muscle-specific elaboration of focal adhesions found in cultured nonmuscle cells (35). As techniques were developed to isolate and maintain neonatal and adult cardiomyocytes in long-term culture, it became apparent that cardiomyocytes remodel their adhesive structures to adapt to the two-dimensional culture environment (49,114). This adaptation includes the reorganization of costameric structures as the cardiomyocytes attach and spread over their ECM-coated substratum, yielding adhesive structures that are structurally similar to typical focal adhesions found in nonmuscle cells (27).…”

Section: Focal Adhesion Complexes In Cultured Cardiomyocytesmentioning

confidence: 99%

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Costameres, focal adhesions, and cardiomyocyte mechanotransduction

Samarel¹

2005

American Journal of Physiology-Heart and Circulatory Physiology

252

201

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Samarel, Allen M. Costameres, focal adhesions, and cardiomyocyte mechanotransduction. Am J Physiol Heart Circ Physiol 289: H2291-H2301, 2005; doi: 10.1152/ajpheart.00749.2005.-Mechanotransduction refers to the cellular mechanisms by which load-bearing cells sense physical forces, transduce the forces into biochemical signals, and generate appropriate responses leading to alterations in cellular structure and function. This process affects the beat-to-beat regulation of cardiac performance but also affects the proliferation, differentiation, growth, and survival of the cellular components that comprise the human myocardium. This review focuses on the experimental evidence indicating that the costamere and its structurally related structure the focal adhesion complex are critical cytoskeletal elements involved in cardiomyocyte mechanotransduction. Biochemical signals originating from the extracellular matrix-integrin-costameric protein complex share many common features with those signals generated by growth factor receptors. The roles of key regulatory kinases and other muscle-specific proteins involved in mechanotransduction and growth factor signaling are discussed, and issues requiring further study in this field are outlined.focal adhesion kinase; proline-rich tyrosine kinase 2; integrin-linked kinase; protein kinase C; signal transduction; heart THE PROCESS OF MECHANOTRANSDUCTION refers to the cellular mechanisms by which load-bearing cells sense physical forces, transduce the forces into biochemical signals, and generate appropriate responses leading to alterations in cellular structure and function. Physical forces encountered by living cells include membrane stretch, gain and loss of adhesion, and compression due to an increase in pressure. The signal transduction pathways that are activated in response to mechanical forces include many unique components, as well as elements shared by other signaling pathways. Mechanotransduction in cardiomyocytes is particularly complex, in that individual muscle cells both respond to externally applied mechanical forces as well as generate internal loads that are transmitted to adjacent cells and their surrounding extracellular matrix (ECM). Mechanotransduction in both atrial and ventricular cardiomyocytes affects the beat-to-beat regulation of cardiac performance but also profoundly affects the proliferation, differentiation, growth, and survival of the cellular components that comprise the human myocardium. Understanding the cellular and molecular basis for mechanotransduction is therefore important to our overall understanding of cardiac structure and function in the normal and diseased heart.Observational studies conducted during the 1970s addressing the hypertrophic growth response of human myocardium to pathological changes in systolic and diastolic wall stress (42) fostered the development of experimental model systems with which to explore cardiomyocyte mechanotransduction in vivo and in vitro. These model systems now span the breadth of experimental cardiology...

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“…Cell cultures of cardiomyocytes are commonly used in vitro model for investigation at a cellular level. Primary cardiomyocytes have been successfully isolated from hearts of a range of species [e.g., mouse (16), rat (4), guinea pig (15), rabbit (12), cat (31), and human (27)] using a simple enzymatic method and maintained as primary cultures for a short duration (13).…”

mentioning

confidence: 99%

Enrichment of neonatal rat cardiomyocytes in primary culture facilitates long-term maintenance of contractility in vitro

Nguyen

Hsiao

Sivakumaran

et al. 2012

American Journal of Physiology-Cell Physiology

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Enrichment of neonatal rat cardiomyocytes in primary culture facilitates long-term maintenance of contractility in vitro. Am J Physiol Cell Physiol 303: C1220 -C1228, 2012. First published August 29, 2012; doi:10.1152/ajpcell.00449.2011.-Long-term culture of primary neonatal rat cardiomyocytes is limited by the loss of spontaneous contractile phenotype within weeks in culture. This may be due to loss of contractile cardiomyocytes from the culture or overgrowth of the non-cardiomyocyte population. Using the mitochondria specific fluorescent dye, tetramethylrhodamine methyl ester perchlorate (TMRM), we showed that neonatal rat cardiomyocytes enriched by fluorescence-activated cell sorting can be maintained as contractile cultures for long periods (24-wk culture vs. 2 wk for unsorted cardiomyocytes). Long-term culture of this purified cardiomyocyte (TMRM high) population retained the expression of cardiomyocyte markers, continued calcium cycling, and displayed cyclic electrical activity that could be regulated pharmacologically. These findings suggest that non-cardiomyocyte populations can negatively influence contractility of cardiomyocytes in culture and that by purifying cardiomyocytes, the cultures retain potential as an experimental model for longitudinal studies of cardiomyocyte biology in vitro.cardiomyocyte; tetramethylrhodamine methyl ester perchlorate; enrichment; contractility CARDIOVASCULAR DISEASE is a leading cause of morbidity and mortality in the world (21). The ongoing pandemic of cardiovascular disease has fueled much basic science research to achieve a better understanding of the pathophysiological function of cardiomyocytes (CMs) and for effective strategizing of potential therapies. Cell cultures of cardiomyocytes are commonly used in vitro model for investigation at a cellular level. Primary cardiomyocytes have been successfully isolated from hearts of a range of species [e.g., mouse (16), rat (4), guinea pig (15), rabbit (12), cat (31), and human (27)] using a simple enzymatic method and maintained as primary cultures for a short duration (13).Neonatal rat CM cultures have been widely employed as a cost-effective experimental model to study the contractility, electrophysiology, and morphology of CMs. CMs from postnatal hearts do not normally proliferate in culture (30), thus limiting the number of CMs available to the starting material. Long-term primary cultures of CMs are rarely reported in the literature; generally experimental time frames are limited to 2-3 wk. Previous studies have shown that neonatal CMs lose their contractile phenotype within weeks in culture and that this is associated with CM dedifferentiation and cytoskeleton remodeling (3, 26). One potential reason for this could be the cellular heterogeneity of the heart, containing both contractile CMs and noncontractile cells such as fibroblasts, endothelial cells, and smooth muscle cells. The noncontractile cells, especially fibroblasts, can proliferate rapidly in culture and typically overgrow CMs to a point where it is belie...

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Contractile activity and cell-cell contact regulate myofibrillar organization in cultured cardiac myocytes.

Cited by 80 publications

References 47 publications

C-Kit Positive Cells from Failing Human Hearts: Role of Culturing Media on Cardiomyogenic Potentials

C-Kit Positive Cells from Failing Human Hearts: Role of Culturing Media on Cardiomyogenic Potentials

Costameres, focal adhesions, and cardiomyocyte mechanotransduction

Enrichment of neonatal rat cardiomyocytes in primary culture facilitates long-term maintenance of contractility in vitro

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