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  • 11
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    Improved Tissue Culture Conditions for Engineered Skeletal Muscle Sheets (2013)
    Hindawi
    Details
    Publication Date: 2013-01-01
    Description: The potential clinical utility of engineered muscle is currently restricted by limitedin vitrocapacity of expanded muscle precursor cells to fuse and form mature myofibers. The purpose of this study was to use isotropic skeletal muscle sheets to explore the impact of (1) fibroblast coculture and (2) fibroblast-conditioned media (fCM) onin vitromyogenesis. Muscle sheets were prepared by seeding varying ratios of skeletal myoblasts and fibroblasts on a biomimetic substrate and culturing the resulting tissue in either control media or fCM. Muscle sheets were prepared from two cell subpopulations, (1) C2C12 and NOR-10 and (2) primary neonatal rat skeletal muscle cells (nSKM). In C2C12/Nor-10 muscle sheets fCM conferred a myogenic advantage early in culture; at D1 a statistically significant 3.12 ± 0.8-fold increase in myofiber density was observed with fCM. A high purity satellite cell population was collected from an initially mixed population of nSKMs via cell sorting for positiveα7-integrin expression. On D6, tissue sheets with low fibroblast concentrations (0 & 10%) cultured in fCM had increased average myofiber density (4.8 ± 0.2 myofibers/field) compared to tissue sheets with high fibroblast concentrations (50%) cultured in control media (1.0 ± 0.1 myofibers/field). Additionally, fCM promoted longer, thicker myofibers with a mature phenotype.
    Print ISSN: 2356-6140
    Electronic ISSN: 1537-744X
    Topics: Natural Sciences in General
    Published by Hindawi
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    PAPER CURRENT
  • 12
    Electronic Resource
    Electronic Resource
    A Potential Role for Mechanical Stimulation in Cardiac Development (1990)
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 588 (1990), S. 0 
    Details
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1749-6632.1990.tb13196.x
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    Paper (German National Licenses)
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  • 13
    Electronic Resource
    Electronic Resource
    Potential Role of the Extracellular Matrix in Postseptation Development of the Heart (1990)
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 588 (1990), S. 0 
    Details
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1749-6632.1990.tb13199.x
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    Paper (German National Licenses)
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  • 14
    Electronic Resource
    Electronic Resource
    Hormonal Regulation of Polyamine Biosynthetic Enzymes in the Leydig Cell-Derived TM3 Cell Line (1987)
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 513 (1987), S. 0 
    Details
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1749-6632.1987.tb25044.x
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    Paper (German National Licenses)
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  • 15
    Electronic Resource
    Electronic Resource
    Myofibrillar and cytoskeletal assembly in neonatal rat cardiac myocytes cultured on laminin and collagen (1991)
    Springer
    Cell & tissue research 264 (1991), S. 577-587 
    Details
    ISSN: 1432-0878
    Keywords: Myofibrils ; Cytoskeleton ; Extracellular matrix ; Laminin ; Collage ; Actin filaments ; Vinculin-Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neonatal rat cardiomyocytes were cultured on extracellular matrix components laminin and collagens I+III to examine effects of extracellular matrix on the assembly of cytoskeletal proteins during myofibrillogenesis. Myofibril assembly was visualized by immunofluorescence of marker proteins for myofibrils (f-actin for I bands and α-actinin for Z bands), focal adhesions (vinculin), and transmembrane extracellular matrix receptors (β1 integrin) as cells spread for various times in culture. By 4 h in culture, f-actin appeared organized into nonstriated stress-fiber-like structures while α-actinin, vinculin and β1 integrin were localized in small streaks and beads. Subsequently, striated patterns were observed sequentially in the intracellular cytoskeletal components α-actinin, vinculin, f-actin, and then in the transmembrane β1 integrin receptor. These data support an earlier model for sarcomerogenesis in which stress-fiber-like structures serve as initial scaffolds upon which α-actinin and then vinculin-containing costameres are assembled. This sequential and temporal assembly was the same on both laminin and collagens I+III. A quantitative difference, however, was apparent on the 2 matrices. There was an increased appearance on collagens I+III of rosettes (also called podosomes or cortical actin-containing bodies in other cells) which consisted of an f-actin core surrounded by α-actinin, vinculin and β1 integrin rims. The increased incidence of rosettes in neonatal myocytes on collagens I+III suggests that these cytoskeletal complexes are involved in recognition and interaction with extracellular matrix components.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00319047
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  • 16
    Electronic Resource
    Electronic Resource
    A densitometer for the evaluation of cell growth in primary cultures: Construction and operation (1982)
    Springer
    Methods in cell science 7 (1982), S. 5-8 
    Details
    ISSN: 1573-0603
    Keywords: photovoltic cell densitometer ; primary cell culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This report describes the design, construction and operation of a photovoltic cell densitometer. The densitometer has proven to be very useful in evaluating the effect of various culture medium supplements on the growth of primary cultures.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01666872
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  • 17
    Electronic Resource
    Electronic Resource
    Primary culture of canine prostate epithelial cells (1982)
    Springer
    Methods in cell science 7 (1982), S. 51-54 
    Details
    ISSN: 1573-0603
    Keywords: prostate ; canine ; primary culture ; testosterone metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Epithelial-cell enriched primary cultures were established from canine prostate by a collagenase digestion and selective attachment procedure. The epithelial cells in primary culture retained differentiated structure and function. The epithelial cells were attached to one another by tight junctions and desmosomes to form “lumenlike structures” that resemble the acini of the intact organ and appeared to contain typical protein synthetic organelles (1,2). The cultures contained significant levels of acid phosphatase and ornithine decarboxylase (2) and retained the ability to metabolize testosterone to dihydrotestosterone and other 5α-reduced metabolites (2–4).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01665908
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  • 18
    Electronic Resource
    Electronic Resource
    Primary culture of rat ventral prostate epithelial cells (1979)
    Springer
    Methods in cell science 5 (1979), S. 1169-1171 
    Details
    ISSN: 1573-0603
    Keywords: rat ventral prostate ; primary culture ; epithelial cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00920580
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  • 19
    Electronic Resource
    Electronic Resource
    The role of β1 integrin in spreading and myofibrillogenesis in neonatal rat cardiomyocytes in vitro (1992)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 21 (1992), S. 87-100 
    Details
    ISSN: 0886-1544
    Keywords: myofibrils ; extracellular matrix ; cytoskeleton ; integrins ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The influence of the extracellular matrix (ECM) on cell behavior, myofibrillogenesis and cytoarchitecture was investigated in neonatal rat cardiac myocytes in vitro. Cell behavior was examined by analyzing cell spreading on different ECM components under a variety of experimental conditions. Area measurements were made on digitized images of cells grown for various time intervals on fibronectin (FN), laminin (LN), collagens I and III (C I + III), plastic, and bovine serum albumin (BSA). The amount of spreading was varied on the different matrices and was maximal on FN 〉 LN 〉 C I+III 〉 plastic 〉 BSA. Addition of anti-β1 integrin antibodies to myocytes cultured on FN, LN and C I+III blocked spreading outward on the substrates and altered normal myofibrillogenesis, especially on LN. Concomitantly, the integrin antibodies induced the formation of giant pseudopodial processes which protruded upward from the substrates. These pseudopods contained actin polygonal networks which exhibited a regular geometrical configuration.Effects of the ECM on cytoarchitecture was examined by analyzing the temporal and spatial patterns of fluorescence and immunogold labeling of cytoskeletal and integrin proteins as myocytes spread in culture. The first indication of sarcomeric patterns was the appearance at 4 hours of striations formed by lateral alignment of α-actinin aggregates into Z bands. At later times, vinculin at 8 hours and β integrin at 22 hours became co-localized with α-actinin at the Z bands and focal adhesions. These data indicate that ECM components influence myocyte spreading and that myofibril assembly and/or stability is associated with ECM-integrin-cytoskeleton associations.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cm.970210202
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  • 20
    Electronic Resource
    Electronic Resource
    In vitro studies on adult cardiac myocytes: Attachment and biosynthesis of collagen type IV and laminin (1988)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 136 (1988), S. 43-53 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The interactions between adult rat cardiac myocytes and the basement membrane components collagen type IV and laminin were investigated in attachment experiments and biosynthesis studies and by immunofluorescence staining. Adult myocytes attached equally well to native collagen type IV and laminin but did not attach to collagen type IV solubilized with pepsin (P-CIV) or to collagen type I. However, when laminin was used to coat P-CIV, attachment was enhanced. Affinity-purified antibodies against laminin inhibited the attachment of myocytes to dishes coated with native collagen type IV, indicating that cell surface-bound laminin mediated attachment of the cells to this substrate. Immunofluorescence staining of freshly isolated myocytes, using antibodies against laminin or collagen type IV, revealed the presence of laminin but not of collagen type IV on the surface of freshly isolated cells, indicating that during the isolation procedure collagen IV was removed from the cell surface. Metabolic labeling followed by immunoprecipitation demonstrated synthesis of both laminin and collagen type IV in cardiac myocytes as they progressed into culture over a 14-day period. This synthesis was accompanied by the deposition of the collagen type IV and laminin into distinctly different patterns as revealed by immunofluorescence staining. As the cells progressed into culture, newly synthesized laminin formed a network radiating from the center of the reorganizing cell into the pseudopods. The laminin was redistributed and remodeled with time in culture to form a dense layer beneath the cell. Collagen type IV was also synthesized with time in culture, but the pattern was a much finer network as opposed to the denser pattern of laminin staining. These studies demonstrate that adult cardiac myocytes synthesize and remodel the basement membrane as they adapt to the culture environment.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041360106
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