Long-term culture of muscle explants from Sparus aurata

Funkenstein, Bruria; Balas, Valentina E.; Skopal, T.; Radaelli, Giuseppe; Rowlerson, A

doi:10.1016/j.tice.2006.09.003

Cited by 25 publications

(15 citation statements)

References 55 publications

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“…In particular, proliferation and differentiation processes may have occurred in the muscle explants, as shown by the differential expression of pcna and myog. These observations are corroborated by a previous ex vivo study in sea bream (Sparus aurata) muscle explants [24]. Given the above features, both the myosatellite cell culture and muscle explants can be used as models to investigate the autonomy of some centralized biological processes in skeletal muscle, such as the circadian rhythm.…”

Section: Discussionsupporting

confidence: 77%

“…On the other hand, muscle explants can demonstrate how the muscle continues to function as a tissue unit, rather than just a cell type. Therefore, muscle explants do not only provide an approximation of the in vivo conditions for cell proliferation and differentiation but also enable a closer comparison with muscle regeneration events in vivo [24]. The differential regulation of some myogenic markers within 5 days of culture of the muscle explants indicates that myogenic activity of the muscle was sustained under ex vivo conditions.…”

Section: Discussionmentioning

confidence: 99%

“…Muscle explants were obtained following the method of Funkenstein et al [24], with minor modifications. After the fish were euthanatized, a fast skeletal muscle fragment from the …”

Section: Short-term Culture Of Fast Skeletal Muscle Explants Of Atlanmentioning

confidence: 99%

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In vitro and ex vivo models indicate that the molecular clock in fast skeletal muscle of Atlantic cod is not autonomous

et al. 2014

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Section: Discussionsupporting

confidence: 77%

Section: Discussionmentioning

confidence: 99%

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In vitro and ex vivo models indicate that the molecular clock in fast skeletal muscle of Atlantic cod is not autonomous

et al. 2014

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“…Studies undertaken by Perrot et al (1999Perrot et al ( , 2000 have shown that IGF-IRs and their ligands (IGF-I and IGF-II) are expressed during early development stages and gonad development in the gilthead sea bream. Recently, Funkenstein et al (2006) have reported the long-term culture of muscle explants from gilthead sea bream and found that the new muscle fibres express IGFs by 1 week and IGF-IRs by 23 days.…”

Section: Introductionmentioning

confidence: 99%

IGF-I binding and receptor signal transduction in primary cell culture of muscle cells of gilthead sea bream: changes throughout in vitro development

et al. 2007

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We examined the possibility of culturing muscle cells of gilthead sea bream in vitro and assessed variations in insulin-like growth factor-I (IGF-I) binding during myocyte development. The viability of the cell culture was determined by fluorescence-activated cell-sorting analysis, which showed that the percentage of dead cells decreased with cell differentiation. The intracellular reduction of MTT into formazan pigment was preferentially carried out as cells differentiated (from day 4) indicating an increase in metabolic activity. IGF-I-binding assays demonstrated that the number of receptors increased from 190 +/- 0.09 fmol/mg protein in myocytes at day 5 to 360 +/- 0.09 fmol/mg protein in myotubes at day 12. The affinity of IGF-I receptors did not change significantly during cell development (from 0.89 +/- 0.09 to 0.98 +/- 0.09 nM). The activation of various kinase (ERK 1/2 MAPK and Akt/PKB) proteins by IGFs and insulin was studied by means of Western blot analysis. Levels of MAPK-P increased after IGF and insulin treatment during the first stages of cell culture, with a low response being observed at day 15, whereas IGFs displayed a stimulatory effect on Akt-P throughout the cell culture period, even on day 15. This study thus shows that (1) gilthead sea bream myocytes can be cultured, (2) they express functional IGF-I receptors that increase in number as they differentiate in vitro; (3) IGF signalling transduction through IGF-I receptors stimulates the MAPK and Akt pathways, depending on the development stage of the muscle cell culture.

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“…The lack of dark staining of the larger cell type by anti-MHC suggests that these cells might be another population of cells that is not as fully differentiated as the MHC+ cells. Working with satellite cells from rainbow trout and yellow perch has proved to be challenging (Funkenstein et al 2006). The quantities of satellite cells isolated and the proliferation rates of these cells are lower than those we have obtained in other animal species.…”

Section: Discussionmentioning

confidence: 71%

Unidentified cells reside in fish skeletal muscle

et al. 2008

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Cell cultures were established from the skeletal muscle tissue of 6-13 months old rainbow trout and 12-14 months old yellow perch. Approximately 27,000 ± 5,000 cells/g (trout; N = 5) and 5,000 ± 1,200 cells/g of tissue (perch; N = 4) were obtained. Isolation and propagation were qualitatively greater for both species when the cells (younger fish producer more cells than older fish) were exposed to DMEM + 15% FBS, rather than L-15 + 15% FBS, at 20°C (trout) and at 24°C (yellow perch). Two morphologically distinct cell types were observed in cultures of both species, some of which eventually formed very small myotubes, which displayed immunocytological reactivity for myogenin, myosin heavy chain, and a-actinin; the second population of cells remained unstained. Successful cryopreservation was achieved using a 5% DMSO and 95% serum mixture, but post-thawing viabilities were low 5-27% (trout) and 14-30% (perch). Further research is needed in order to determine cell type specificity of isolated cells.

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Long-term culture of muscle explants from Sparus aurata

Cited by 25 publications

References 55 publications

In vitro and ex vivo models indicate that the molecular clock in fast skeletal muscle of Atlantic cod is not autonomous

In vitro and ex vivo models indicate that the molecular clock in fast skeletal muscle of Atlantic cod is not autonomous

IGF-I binding and receptor signal transduction in primary cell culture of muscle cells of gilthead sea bream: changes throughout in vitro development

Unidentified cells reside in fish skeletal muscle

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