Temporal expression of growth factor genes of primary porcine satellite cells during myogenesis

Theil, Peter Kappel; Sørensen, I; Nissen, P. M.; Oksbjerg, Niels

doi:10.1111/j.1740-0929.2006.00356.x

Cited by 33 publications

(29 citation statements)

References 30 publications

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“…Satellite cell isolation procedure The method for isolation of SCs has previously been described for postnatal and foetal muscle tissue in pigs (Nissen et al, 2005;Theil et al, 2006). In short, the LW, MW and HW female pigs within each of the eight litters were stunned by the CO 2 method.…”

Section: Animalsmentioning

confidence: 99%

In vitro primary satellite cell growth and differentiation within litters of pigs

Nissen

Oksbjerg

2009

Animal

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Postnatal muscle growth is dependent on satellite cell (SC) proliferation, differentiation and fusion to increase the DNA content of existing muscle fibres and thereby the capacity to synthesize protein. The purpose of the present study was to examine the ability of isolated SCs from low, medium and high weaning weight litter mates of pigs to proliferate and differentiate, and to affect protein synthesis and degradation after fusion into myotubes. At 6 weeks of age, SCs from the lowest weight (LW), medium weight (MW) and highest weight (HW) female pigs within eight litters were isolated. Thereby, eight cultures of SCs were established for each of the three weight groups within litter, representing three groups of SCs from pigs exhibiting differences in postnatal muscle growth performance. Proliferation was estimated as the number of viable cells at different time points after seeding. SC differentiation was evaluated by measuring the activity of the muscle-specific enzyme, creatine phosphokinase, and protein synthesis and degradation were measured by incorporation and release of 3 H-tyrosine, respectively. A tendency towards a difference in proliferation between SC cultures was found ( P 5 0.09). This was evident as the number of viable cells at day 3 was lower in cultures from LW pigs than from HW ( P , 0.05) and MW ( P , 0.01) pigs. Differentiation was significantly different between cultures ( P , 0.05). There was a significant difference between LW and MW cultures at 72 h ( P , 0.05), and a tendency towards a difference between LW and HW cultures at 45 h ( P 5 0.07). Protein synthesis per mg protein or per mg DNA did not differ among SC cultures from LW, MW and HW pigs. Neither did protein degradation rate differ significantly among SC cultures from LW, MW and HW pigs. Overall, the results show that SCs from LW pigs seem to proliferate and differentiate at a slower rate than SCs from MW and HW pigs. The results found in this study show no difference in the ability of SCs to affect protein synthesis or degradation between SCs from litter mates exhibiting different growth rates in vivo.

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Section: Animalsmentioning

confidence: 99%

In vitro primary satellite cell growth and differentiation within litters of pigs

Nissen

Oksbjerg

2009

Animal

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“…The original method of Bischoff was used with some modifications (Bischoff 1974;Theil et al 2006). Muscle tissue was excised, stripped for visible fat and connective tissue, placed in ice-cold transport medium (1% glucose, 500 IU/ml of penicillin, 500 μg/ml of streptomycin sulfate, 15 μg/ml of amphotericin and 100 μg/ml of gentamycin in Ca 2+ and Mg 2+ free phosphate buffered saline (PBS)), and transferred to a laminar flow bench.…”

Section: Porcine Primary Myoblast and Myotube Culturesmentioning

confidence: 99%

The Polyacetylenes Falcarinol and Falcarindiol Affect Stress Responses in Myotube Cultures in a Biphasic Manner

et al. 2008

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h The effects of the bioactive polyacetylenes, falcarinol and falcarindiol, present in carrots, celery, celeriac and other umbelliferous vegetables, on the stress responses in primary myotube cultures, were studied. Biphasic responses on cellular stress responses in myotube cultures were investigated by exposing them to various concentrations of falcarinol and falcarindiol for 24 h before testing effects of 100 μM H 2 O 2 on the intracellular formation of reactive oxygen species (ROS), transcription of the antioxidative enzyme cytosolic glutathione peroxidase (cGPx), and the heat shock proteins (HSP) HSP70 and HO1. At low concentrations (1.6 to 25 μM) polyacetylenes caused a slightly accelerated intracellular ROS formation, increased cGPx transcription and decreased HSP70 and HO1 transcription. The increased cGPx transcription may be interpreted as an adaptive response to the increased ROS formation and may have caused a reduced demand for the protective functions of the HSPs. ROS formation, however, was substantially decreased after pre-incubation with both polyacetylenes at 50 and 100 μM, the cGPx transcription was reduced and the HSP70 and HO1 transcription increased, indicating a need for the protective and repairing functions of the HSPs. In conclusion, pre-incubation with low concentrations of both polyacetylenes prior to H 2 O 2 exposure induced a cytoprotective effect whereas higher concentrations had adverse effects.

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“…C q values of technical replicates were averaged then used to determine ΔC q values (ΔC q = C q of the target gene/miRNA − C q of the reference gene/miRNA) (Theil et al, 2006a). The 2…”

Section: Discussionmentioning

confidence: 99%

Temporal correlation between differentiation factor expression and microRNAs in Holstein bovine skeletal muscle

Miretti

Volpe

Martignani

et al. 2017

Animal

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Satellite cells are adult stem cells located between the basal lamina and sarcolemma of muscle fibers. Under physiological conditions, satellite cells are quiescent, but they maintain a strong proliferative potential and propensity to differentiate, which underlies their critical role in muscle preservation and growth. MicroRNAs (miRNAs) play essential roles during animal development as well as in stem cell self-renewal and differentiation regulation. MiRNA-1, miRNA-133a and miRNA-206 are closely related muscle-specific miRNAs, and are thus defined myomiRNAs. MyomiRNAs are integrated into myogenic regulatory networks. Their expression is under the transcriptional and post-transcriptional control of myogenic factors and, in turn, they exhibit widespread control of muscle gene expression. Very little information is available about the regulation and behavior of satellite cells in large farm animals, in particular during satellite cell differentiation. Here, we study bovine satellite cells (BoSCs) undergoing a differentiation process and report the expression pattern of selected genes and miRNAs involved. Muscle samples of longissimus thoracis from Holstein adult male animals were selected for the collection of satellite cells. All satellite cell preparations demonstrated myotube differentiation. To characterize the dynamics of several transcription factors expressed in BoSCs, we performed real-time PCR on complementary DNA generated from the total RNA extracted from BoSCs cultivated in growth medium (GM) or in differentiation medium (DM) for 4 days. In the GM condition, BoSCs expressed the satellite cell lineage markers as well as transcripts for the myogenic regulatory factors. At the time of isolation from muscle, PAX7 was expressed in nearly 100% of BoSCs; however, its messenger RNA (mRNA) levels dramatically decreased between 3 and 6 days post isolation ( P < 0.01). MyoD mRNA levels increased during the 1st day of cultivation in DM (day 7; P < 0.02), showing a gradual activation of the myogenic gene program. During the subsequent 4 days of culture in DM, several tested genes, including MRF4, MYOG, MEF2C, TMEM8C, DES and MYH1, showed increased expression ( P < 0.05), and these levels remained high throughout the culture period investigated. Meanwhile, the expression of genes involved in the differentiation process also miRNA-1, miRNA-133a and miRNA-206 were strongly up-regulated on the 1st day in DM (day 7; P < 0.05). Analysis revealed highly significant correlations between myomiRNAs expression and MEF2C, MRF4, TMEM8C, DES and MYH1 gene expression ( P < 0.001). Knowledge about the transcriptional changes correlating with the growth and differentiation of skeletal muscle fibers could be helpful for developing strategies to improve production performance in livestock.

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Temporal expression of growth factor genes of primary porcine satellite cells during myogenesis

Cited by 33 publications

References 30 publications

In vitro primary satellite cell growth and differentiation within litters of pigs

In vitro primary satellite cell growth and differentiation within litters of pigs

The Polyacetylenes Falcarinol and Falcarindiol Affect Stress Responses in Myotube Cultures in a Biphasic Manner

Temporal correlation between differentiation factor expression and microRNAs in Holstein bovine skeletal muscle

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