記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： https://doi.org/10.1515/ijfe-2024-0255

記述言語：英語   出版者・発行元：Biotechnology Journal  

Skeletal muscle satellite cells (SCs) are essential for muscle regeneration. Their proliferation and differentiation are influenced by fibroblast growth factor (FGF)-2. In this study, we screened for FGF-2-derived peptides that promote SC proliferation. Utilizing photocleavable peptide array technology, a library of 7-residue peptides was synthesized, and its effect on SC proliferation was examined using a mixture of five peptides. The results showed that peptides 1–5 (136%), 21–25 (136%), 26–30 (141%), 31–35 (159%), 71–75 (135%), 76–80 (144%), and 126–130 (137%) significantly increased SC proliferation. Further experiments revealed that peptide 33, CKNGGFF, enhanced SC proliferation. Furthermore, its extended form, peptide 33-13, CKNGGFFLRIHPD, promoted SC proliferation and increased the percentage of Pax7-positive cells, indicating that SCs were maintained in an undifferentiated state. The addition of FGF-2 and peptide 33-13 further induced cell proliferation but did not increase the percentage of Pax7-positive cells. A proliferation assay using an FGF receptor (FGFR) inhibitor suggested that peptide 33-13 acts through the FGFR-mediated and other pathways. Although further research is necessary to explore the mechanisms of action of these peptides and their potential for in vivo and in vitro use, the high sequence conservation of peptides 33 and 33-13 in FGF-2 across multiple species suggests their broad application prospects in biomedical engineering and biotechnology.

DOI： 10.1002/biot.202400278

Open Access

Web of Science

Scopus

PubMed

記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Biochemical Engineering Journal  

Bile acid (BA) micelle-disrupting peptides are bioactive peptides (BPs) that suppress intestinal cholesterol adsorption and ameliorate blood cholesterol levels. To facilitate their application in the food industry, an effective and straightforward technology for BP separation and enrichment of BPs is required. For this purpose, we assessed the potential of heat-treated (HT) porous silica gels with an enhanced capacity for peptide adsorption/desorption (AD). Given the key characteristics of these peptides involving basic amino acid (AA) residues, we applied casein hydrolysate to HT silica gels for adsorption at neutral pH, where the surface yielded anionic silanolates through deprotonation. Subsequently, desorption was conducted at pH 2, at which point the surface returned to its neutral state. We confirmed a 335-fold increase in the BA micelle-disrupting activity of the resulting hydrolysates. LC-MS/MS analysis identified 783 peptides, confirming that the peptides containing cationic AA residues were selectively enriched, as expected. Moreover, a dose-response of randomly selected 27 peptides with various enrichment levels revealed the presence of multiple active peptides in the enriched groups. These results indicate that our method is effective for the separation and enrichment of BA micelle-disrupting peptides and holds promise for BP production.

DOI： 10.1016/j.bej.2024.109283

Web of Science

Scopus

記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Biochemical and Biophysical Research Communications  

Extracellular vesicles (EVs) released into the blood during exercise mediate its whole-body health effects. The differentiation of EVs released by skeletal muscle cells in vivo from those released by other cells is challenging, therefore, it is unclear whether exercise increases the number of EVs secreted by skeletal muscle cells. In this study, we investigated whether exercise affects the quantity of EVs released from skeletal muscle cells using in vitro exercise models. C2C12 myotubes were cultured on a gel layer with 1 or 30 Hz electrical pulse stimulation (EPS) to induce contractions as an artificial simulating exercise. We found that tetanic contraction induced by 30 Hz EPS increased the number of secreted EVs. MicroRNA (miRNA)-seq analysis revealed that 30 Hz EPS altered the miRNA in the secreted EVs. Furthermore, expression analysis of genes related to the biogenesis and transport of EVs revealed that the expression of ALG-2 interacting protein X (Alix) was increased in response to 30 Hz EPS, and the peak value of intracellular Ca2+ in myotubes at 30 Hz EPS was higher than that at 1 Hz, indicating that the increase in intracellular Ca2+ concentration may be related to the increased secretion of EVs in response to 30 Hz EPS.

DOI： 10.1016/j.bbrc.2023.06.054

Web of Science

Scopus

PubMed

記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Cells  

In vitro neuromuscular junction (NMJ) models are powerful tools for studying neuromuscular disorders. Although linearly patterned culture surfaces have been reported to be useful for the formation of in vitro NMJ models using mouse motor neuron (MNs) and skeletal muscle (SkM) myotubes, it is unclear how the linearly patterned culture surface increases acetylcholine receptor (AChR) clustering, one of the steps in the process of NMJ formation, and whether this increases the in vitro NMJ formation efficiency of co-cultured human MNs and SkM myotubes. In this study, we investigated the effects of a linearly patterned culture surface on AChR clustering in myotubes and examined the possible mechanism of the increase in AChR clustering using gene expression analysis, as well as the effects of the patterned surface on the efficiency of NMJ formation between co-cultured human SkM myotubes and human iPSC-derived MNs. Our results suggest that better differentiation of myotubes on the patterned surface, compared to the flat surface, induced gene expression of integrin α7 and AChR ε-subunit, thereby increasing AChR clustering. Furthermore, we found that the number of NMJs between human SkM cells and MNs increased upon co-culture on the linearly patterned surface, suggesting the usefulness of the patterned surface for creating in vitro human NMJ models.

DOI： 10.3390/cells11233760

Open Access

Web of Science

Scopus

PubMed

種別：学会・研究会等 

種別：学会・研究会等