Growth and differentiation of permanent and secondary mouse myogenic cell lines on microcarriers

Bardouille, Claudia; Lehmann, J.; Heimann, Peter; Jockusch, Harald

doi:10.1007/s002530100595

Cited by 41 publications

(30 citation statements)

References 22 publications

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“…1B). Once they undergo differentiation, the C2C12 myotubes do not continue to divide, but they remain viable for approximately 21 days in vitro, after which time spontaneous contraction can occur, causing detachment from the culture surface (6,20,35).…”

Section: Resultsmentioning

confidence: 99%

Prion Infection of Muscle Cells In Vitro

Dlakic

Grigg

Bessen

2007

J Virol

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The prion agent has been detected in skeletal muscle of humans and animals with prion diseases. Here we report scrapie infection of murine C2C12 myoblasts and myotubes in vitro following coculture with a scrapieinfected murine neuroblastoma (N2A) cell line but not following incubation with a scrapie-infected nonneuronal cell line or a scrapie brain homogenate. Terminal differentiation of scrapie-infected C2C12 myoblasts into myotubes resulted in an increase in the expression of the disease-specific prion protein, PrP Sc . The amount of scrapie infectivity or PrP Sc in C2C12 myotubes was comparable to the levels found in scrapie-infected N2A cells, indicating that a high level of infection was established in muscle cells. Subclones of scrapie-infected C2C12 cells produced high levels of PrP Sc in myotubes, and the C-terminal C2 polypeptide fragment of PrP Sc was found based on deglycosylation and PrP Sc -specific immunoprecipitation of cell lysates. This is the first report of a stable prion infection in muscle cells in vitro and of a long-term prion infection in a nondividing, differentiated peripheral cell type in culture. These in vitro studies also suggest that in vivo prion infection of skeletal muscle requires contact with prion-infected neurons or, possibly, nerve terminals.

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Section: Resultsmentioning

confidence: 99%

Prion Infection of Muscle Cells In Vitro

Dlakic

Grigg

Bessen

2007

J Virol

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“…Inhibition of differentiation by MRP8/MRP14 was reflected by a significant reduction of myotube formation and confirmed by an inhibitory effect on the expression of myocytic differentiation markers such as myogenin. 35,42 In addition, proliferation of C2C12 cells was blocked in the presence of MRP8/MRP14. At higher concentrations MRP8/MRP14 induced characteristic morphological and ultrastructural changes of apoptosis in C2C12 cells as shown by light and electron microscopy.…”

Section: Discussionmentioning

confidence: 99%

Expression of Calcium-Binding Proteins MRP8 and MRP14 in Inflammatory Muscle Diseases

Seeliger

Vogl

Engels

et al. 2003

The American Journal of Pathology

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The pathophysiological role of infiltrating macrophages and their subtypes in idiopathic inflammatory myopathies such as dermatomyositis, polymyositis, and inclusion body myositis is not fully clear. Monocytes exhibit various phenotypes with different functional properties such as release of pro-or anti-inflammatory mediators. Expression of myeloid-related proteins MRP8 and MRP14, two calcium-binding S100-proteins, characterizes a proinflammatory subtype of macrophages. We immunohistochemically investigated expression of MRP8 and MRP14 in muscle biopsies of 33 patients with dermatomyositis, polymyositis, and inclusion body myositis. We found a clear association of expression of MRP8 and MRP14 by infiltrating macrophages with degeneration of myofibers. Because MRP8 and MRP14 are secreted by activated macrophages we investigated if these proteins would have direct extracellular effects on myocytes. We found that the purified MRP8/MRP14 complex inhibited proliferation and differentiation of C2C12 myoblasts and that it induced apoptosis via activation of caspase-3 in a time-and dose-dependent manner. These results indicate that in the course of inflammatory myopathies, activated macrophages can promote destruction and impair regeneration of myocytes via secretion of MRP8/MRP14. Myositis is a term describing muscle inflammation independent of its etiology. The heterogeneous group of acute and chronic idiopathic inflammatory myopathies (IM) is histologically characterized by signs of destruction and partial regeneration of inflamed muscle fibers.

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“…In the spinner flask scaffolds are fixed on needles or seeded onto microcarriers (13)(14)(15). A well-mixed environment is maintained with a magnetic stir bar at the bottom of the flask.…”

Section: Introductionmentioning

confidence: 99%

Design and Characterization of a Rotating Bed System Bioreactor for Tissue Engineering Applications

Anton¹,

Suck²,

Diederichs³

et al. 2008

Biotechnol. Prog.

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The main challenge in the development of bioreactors for tissue engineering is the delivery of a sufficient nutrient and oxygen supply for cell growth in a 3D environment. Thus, a new rotating bed system bioreactor for tissue engineering applications was developed. The system consists of a culture vessel as well as an integrated rotating bed of special porous ceramic discs and a process control unit connected with the reactor to ensure optimal culturing conditions. The aim of the project was the design and construction of a fully equipped rotating bed reactor, and in particular, the characterization and optimization of the system with regard to technical parameters such as mixing time and pH-control to guarantee optimal conditions for cell growth and differentiation. Furthermore, the applicability of the developed system was demonstrated by cultivation of osteoblast precursor cells. The porous structure of the ceramic discs and the external medium circulation loop provide an optimal environment for tissue generation in long-term cultivations. Mass transfer limitations were minimized by the slow rotation, which also provides the cells with sufficient nutrients and oxygen through alternate contact to air and medium. An osteoblast precursor cell line was successfully cultivated in this bioreactor for 28 days.

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Growth and differentiation of permanent and secondary mouse myogenic cell lines on microcarriers

Cited by 41 publications

References 22 publications

Prion Infection of Muscle Cells In Vitro

Prion Infection of Muscle Cells In Vitro

Expression of Calcium-Binding Proteins MRP8 and MRP14 in Inflammatory Muscle Diseases

Design and Characterization of a Rotating Bed System Bioreactor for Tissue Engineering Applications

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