• Journal of Life Science
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• v.22 no.10
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• pp.1316-1323
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• 2012

The present study was conducted to investigate whether myoblasts can be transdifferentiated into adipocytes by ectopic expression of adipogenic transcription factors, including peroxisome proliferator-activated receptor-${\gamma}$ ($PPAR{\gamma}$), CCAAT/enhancer-binding protein-${\alpha}$ (C/$EBP{\alpha}$), sterol regulatory element binding protein-1c (SREBP1c), and Krueppel-like factor 5 (KLF5), in primary bovine satellite cells. Transcription factors were transiently transfected into primary bovine myoblasts, and the cells were cultured with adipogenic differentiation medium for 2 days and then cultured on growth medium for an additional 8 days. Ectopic expression of $PPAR{\gamma}$ or C/$EBP{\alpha}$ alone was insufficient to induce adipogenesis in myoblasts. However, overexpression of both $PPAR{\gamma}$ and C/$EBP{\alpha}$ in myoblasts was able to induce adipogenic transdifferentiation as indicated by the appearance of mature adipocytes, the induction of adipogenic gene expressions, and the suppression of myogenic gene expressions. In addition, KLF5 and $PPAR{\gamma}$ co-transfected bovine myoblasts were converted to adipocytes but not in cells transfected with only KLF5 expression vector. Overexpression of SREBP1c alone was sufficient to induce transdifferentiation from myoblasts into adipocytes. These results demonstrate that primary bovine satellite cells can be transdifferentiated into adipocytes either by single ectopic expression or combined expression of adipogenic transcription factors in a culture system.

• Animal Bioscience
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• v.34 no.6
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• pp.1078-1087
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• 2021

Objective: Skeletal muscle satellite cells (SMSCs) are significant for the growth, regeneration, and maintenance of skeletal muscle after birth. However, currently, few studies have been performed on the isolation, culture and inducing differentiation of goose muscle satellite cells. Previous studies have shown that C1q and tumor necrosis factor-related protein 3 (CTRP3) participated in the process of muscle growth and development, but its role in the goose skeletal muscle development is not yet clear. This study aimed to isolate, culture, and identify the goose SMSCs in vitro. Additionally, to explore the function of CTRP3 in goose SMSCs. Methods: Goose SMSCs were isolated using 0.25% trypsin from leg muscle (LM) of 15 to 20 day fertilized goose eggs. Cell differentiation was induced by transferring the cells to differentiation medium with 2% horse serum and 1% penicillin streptomycin. Immunofluorescence staining of Desmin and Pax7 was used to identify goose SMSCs. Quantitative realtime polymerase chain reaction and western blot were applied to explore developmental expression profile of CTRP3 in LM and the regulation of CTRP3 on myosin heavy chains (MyHC), myogenin (MyoG) expression and Notch signaling pathway related genes expression. Results: The goose SMSCs were successfully isolated and cultured. The expression of Pax7 and Desmin were observed in the isolated cells. The expression of CTRP3 decreased significantly during leg muscle development. Overexpression of CTRP3 could enhance the expression of two myogenic differentiation marker genes, MyHC and MyoG. But knockdown of CTRP3 suppressed their expression. Furthermore, CTRP3 could repress the mRNA level of Notch signaling pathway-related genes, notch receptor 1, notch receptor 2 and hairy/enhancer-of-split related with YRPW motif 1, which previously showed a negative regulation in myoblast differentiation. Conclusion: These findings provide a useful cell model for the future research on goose muscle development and suggest that CTRP3 may play an essential role in skeletal muscle growth of goose.

• Journal of Animal Science and Technology
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• v.51 no.5
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• pp.413-420
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• 2009

The research has been aimed to investigate the effect of different sera including fetal bovine serum (FBS), male bovine serum (MS), female bovine serum (FS), and castrated-male bovine serum (C-MS) on cell proliferation, follicular maturation and ovulation in vitro. Established cell lines and primary cells were cultured in the culture media supplemented with different sera and cells proliferation was observed by cell counting and MTT assay. The results indicated that cell proliferation was significantly different for different serum source. Proliferation of bovine and human myogenic satellite cells was highest in MS. In contrast, proliferation of breast cancer cells and immune cells were the highest in FS and FBS, respectively. There was no difference in the rate of follicular growth, whereas the rate of ovulation was higher in FBS and C-MS. Finally, the wound healing effect and cell proliferation of 3T3-L1 cells showed that wound healing was fastest in FS and cell proliferation was higher in MS. These results suggest the importance of an optimal serum selection in the experiments involving cell culture system, and gender-specific Hanwoo sera may be used as a substitute to FBS.

• Korean Journal of Ichthyology
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• v.26 no.3
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• pp.171-178
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• 2014

Myogenesis is the formation process of multinucleated myofiber with a contractile capacity from muscle satellite cell (MSCs) during life. This process is tightly controlled by several transcription factors such as Pax3 and Pax7 (paired box protein 3 and 7), MEF2C (myocyte enhancer factor 2) and MRFs (myogenic regulatory factors) etc. On the contrary, myostatin (MSTN) is a transforming growth factor-${\beta}$ superfamily, which functions as a negative regulator of skeletal muscle development and growth. Carnosic acid (CA) is a major phenolic component in rosemary (Rosmarinus officinalis) and have been reported various biological activities such as anticancer, antioxidant, antimicrobial and therapeutic agents for amnesia, dementia, alzheimer's disease. This study was confirmed to effects of CA on muscle cell line and muscle tissue alteration of zebrafish by intramuscular injection or feeding methods. $10{\mu}M$ CA showed a non-cytotoxic on myoblast and a complete inhibition effect against myostatin activity on luciferase assay. In intramuscular injection experiment, the total protein and triglyceride amount of $10{\mu}M/kg$ of CA injected group increased by 11% and decreased by 13% compared to these of the no injected group. In histology analysis of muscle tissues by hematoxylin/eosin staining, the number of muscle fiber of $10{\mu}M/kg$ of CA injected group decreased by 29% and fiber area increased 40% compared to these of no injected group. In feeding experiment, the total protein and triglyceride amount no significance difference compared to these of the normal feeding group. In histology analysis, the number of muscle fiber of 1% CA fed group decreased by 35% and fiber area increased 56% compared to these of normal fed group. We identified that CA have an effect on hypertrophy of muscle fiber in adult zebrafish and the results of this study are considered as the basic data that can reveal the mechanisms of muscle formation via gene and protein level analysis.