• Journal of Animal Science and Technology
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• 제63권4호
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• pp.673-680
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• 2021

The purpose of this study was to establish a basic principal procedure for the processing of cultured meat. The first stage involved isolating satellite cells from the desired muscle of an animal using enzymatic digestion (i.e., by using proteases, collagenases, and pronases). The second stage involved culturing the isolated muscle satellite cells in a growth medium containing fetal bovine serum and penicillin/streptomycin with growth factors for an optimal period of time. The second stage involved a basic method for the isolated muscle cells to proliferate while sub-culturing to further induce differentiation in gelatin-coated culture dishes with the general culture medium. The third stage involved the induction of differentiation of muscle satellite cells or formation of myotubes using myogenic medium. Lastly, the fourth stage involved the identification of cell differentiation or myotube formation (myogenesis) using fluorescent dyes. Moreover, the principle of these protocols can be applied to perform primary culture of animal cells. This study will assist beginners with the technical aspects of culturing meat (isolation, cultivation, and differentiation of muscle satellite cells as well as identification of myotube formation for myogenesis).

• 한국축산식품학회지
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• 제44권4호
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• pp.951-965
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• 2024

Lactiplantibacillus plantarum is a valuable potential probiotic species with various proven health-beneficial effects. L. plantarum LM1001 strain was selected among ten strains of L. plantarum based on proteolytic activity on whey proteins. L. plantarum LM1001 produced higher concentrations of total free amino acids and branched-chain amino acids (Ile, Leu, and Val) than other L. plantarum strains. Treatment of C2C12 myotubes with whey protein culture supernatant (1%, 2% and 3%, v/v) using L. plantarum LM1001 significantly increased the expression of myogenic regulatory factors, such as Myf-5, MyoD, and myogenin, reflecting the promotion of myotubes formation (p<0.05). L. plantarum LM1001 displayed β-galactosidase activity but did not produce harmful β-glucuronidase. Thus, the intake of whey protein together with L. plantarum LM1001 has the potential to aid protein digestion and utilization.

• 생명과학회지
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• 제30권2호
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• pp.202-210
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• 2020

최근까지 근력 개선 또는 만성 근육 손실을 유발하는 질병을 치료하기 위해 많은 연구가 있어왔지만, 지금까지 근력 개선 유도를 위해 대표적으로 사용되는 약물은 단백질 동화 스테로이드(AAS)계열의 steroidal androgen들이 주로 이용되어 왔다. AAS는 다양한 근육 및 체중 손실성 치료를 목적으로 처방되고 있으나, 장기간 또는 과다한 복용은 피부질환, 생식 및 내분비 기능 저하, 심혈관 질환 등 다양한 부작용을 초래한다고 잘 알려져 있다. 따라서, 상대적으로 부작용이 적은 근육 신생을 촉진하는 천연물을 탐색하는 연구는 매우 중요하다. 현재까지 천연물의 처리로 인한 근육 신생의 촉진은 대표적으로 세가지 기전의 영향을 받으며, 그 기전으로는 근육조절인자(MRF)의 양성 조절, 단백질 합성 기전의 활성화 및 단백질 분해기전의 억제가 있다. 본 리뷰에서는, 근육 신생 촉진 효과가 보고된 Black ginseng, Plum 등과 같은 식물 추출물 및 Creatine, Catechin 및 여러 지방산 등 천연물 유래 단일물질에 대하여 소개하고, 더 나아가 현재까지 알려진 상기의 천연물질들의 처리에 의한 근육 신생 촉진 기전에 대하여 기전별로 요약하고자 한다.

• 한국동물학회지
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• 제33권1호
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• pp.108-118
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• 1990

혈청을 비롯해서 extracellular matrix에 존재하는 당단백질인 fibronectin은 근세포의 융합과 밀접한 관계가 있는 것으로 알려져 있다. 본 연구실에서는 최근에 근세포가 분화하는 동안에 fibronectin의 수준이 감소되며, 이러한 감소는 fibronectin의 28 kDa fragment에 대한 수용체으 유용성이 감소하는 결과로 밝혀낸 바 있다. 본 연구에서는 근세포으 융합을 억제하는 물질로 알려진 EGTA를 이용하여 근세포의 융합과 28 kDa fragmen receptor의 관계를 검토하여 보았다.EGTA를 처리한 경우 EGTA를 처리하지 않은 근세포에 비해서 fibronectin의 수준과 28 kDa fragmen binding이 훨씬 적게 감소하였으며, 융합이 봉쇄된 근세포에서 EGTA를 제거하여 융합을 재개시키면 fibronectin의 수준과 28 kDa fragmen의 binding이 정상 근세포 수준으로 환원되었다. 이상의 실험 결과로 볼 때 28 kDa fragmen에 대한 수용체의 감소 또는 변화가 근세포의 분화과정에서 일어나는 fibronectin 수준의 감소와 연관성이 있음을 알 수 있다. 한편, 배양액 내에 trypsin을 처리한 경우에는 처리하지 않은 경우에 비해서 28 kDa fragmen의 binding이 현저하게 감소되었고, gangliosides를 처리한 상태에서는 gangliosides의 농도에 정비례해서 28 kDa fragmen의 binding이 감소되었다. 이 밖에 gel overlay technique을 이용하여 28 kDa fragmen가 SDS-PAGE gel에서 분자량이 약 43kDa인 단백질 및 gangliosides와 binding하는 사실을 알 수 있었다. 이러한 실험 결과를 종합하여 볼 때, 근세포의 융합은 28 kDa fragmen에 대한 receptor의 감소와 관계가 있으며, 그 수용체는 gangliosides와 비슷한 당을 가지고 있는 당단백질일 것으로 추정된다.

• Journal of Ginseng Research
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• 제47권6호
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• pp.726-734
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• 2023

Background: Skeletal muscles play a key role in physical activity and energy metabolism. The loss of skeletal muscle mass can cause problems related to metabolism and physical activity. Studies are being conducted to prevent such diseases by increasing the mass and regeneration capacity of muscles. Ginsenoside Rg5 has been reported to exhibit a broad range of pharmacological activities. However, studies on the effects of Rg5 on muscle differentiation and growth are scarce. Methods: To investigate the effects of Rg5 on myogenesis, C2C12 myoblasts were induced to differentiate with Rg5, followed by immunoblotting, immunostaining, and qRT-PCR for myogenic markers and promyogenic signaling (p38MAPK). Immunoprecipitation confirmed that Rg5 increased the interaction between MyoD and E2A via p38MAPK. To investigate the effects of Rg5 on prevention of muscle mass loss, C2C12 myotubes were treated with dexamethasone to induce muscle atrophy. Immunoblotting, immunostaining, and qRT-PCR were performed for myogenic markers, Akt/mTOR signaling for protein synthesis, and atrophy-related genes (Atrogin-1 and MuRF1). Results: Rg5 promoted C2C12 myoblast differentiation through phosphorylation of p38MAPK and MyoD/E2A heterodimerization. Furthermore, Rg5 stimulated C2C12 myotube hypertrophy via phosphorylation of Akt/mTOR. Phosphorylation of Akt induces FoxO3a phosphorylation, which reduces the expression of Atrogin-1 and MuRF1. Conclusion: This study provides an understanding of how Rg5 promotes myogenesis and hypertrophy and prevents dexamethasone-induced muscle atrophy. The study is the first, to the best of our knowledge, to show that Rg5 promotes muscle regeneration and to suggest that Rg5 can be used for therapeutic intervention of muscle weakness and atrophy, including cancer cachexia.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1996년도 한국생물과학협회 국제학술대회
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• pp.146.2-146
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• 1996

No Abstract, See Full Text

• 한국동물학회:학술대회논문집
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• 한국동물학회 1996년도 한국생물과학협회 국제학술대회
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• pp.147.2-147
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• 1996

No Abstract, See Full Text

• 한국진공학회:학술대회논문집
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• 한국진공학회 2015년도 제49회 하계 정기학술대회 초록집
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• pp.158-158
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• 2015

Myoblast are myogenic precursors that proliferate, activate, and differentiate on muscle injury to sustain the regenerative capacity of skeletal muscle; The neuronal isoform of nitric oxide synthase (nNOS, termed also NOS-I) is expressed in normal adult skeletal muscle, suggesting important functions for Nitric oxide (NO) in muscle biology1,2,3. However, the expression and subcellular localization of NO in muscle development and myoblast differentiation are largely unknown. In this study, we examined effects of the nitric oxide generated by a microwave plasma torch, on proliferation/differentiation of rat myoblastic L6 cells. Experimental data pertaining to nitric oxide production are presented in terms of the oxygen input in units of cubic centimetres per minute. The various levels of nitric oxide are observed depending on the flow rate of nitrogen gas, the ratio of oxygen gas, and the microwave power4. In order to evaluate the potential of nitric oxide as an activator of cell differentiation, we applied nitric oxide generated from the microwave plasma torch to L6 skeletal muscles. Differentiation of L6 cells into myotubes was significantly enhanced the differentiation after nitric oxide treatment. Nitric oxide treatment also increase the expression of myogenesis marker proteins and mRNA level, such as myogenin and myosin heavy chain (MHC), as well as cyclic guanosine monophosphate (cGMP), However during the myotube differentiation we found that NO activate oxidative stress signaling erks expression. Therefore, these results establish a role of NO and cGMP in regulating myoblast differentiation and elucidate their mechanism of action, providing a direct link with oxidative stress signalling, which is a key player in myogenesis. Based on these findings, nitric oxide generated by plasma can be used as a possible activator of cell differentiation and tissue regeneration.

• BMB Reports
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• 제48권11호
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• pp.595-596
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• 2015

Skeletal muscle exhibits a loss of muscle mass and function with age. Decreased regenerative potential of muscle stem/progenitor cells is a major underlying cause of sarcopenia. We analyzed microRNAs (miRNA) that are differentially expressed in young and old myoblasts, to identify novel intrinsic factors that play a degenerative role in aged skeletal muscle. miR-431, one of decreasing miRNAs in old myoblasts, improved the myogenic differentiation when overexpressed in old myoblast, but suppressed their myogenic capability in knockdowned young myoblasts. We found that miR-431 directly binds to 3` untranslated regions (UTR) of Smad4 mRNA, and decreases its expression. Given that SMAD4 is one of the downstream effectors of TGF-β, a well-known degenerative signaling pathway in myogenesis, the decreased miR-431 in old myoblast causes SMAD4 elevation, thus resulting in defective myogenesis. Exogenous expression of miR-431 greatly improved the muscle regeneration in the cardiotoxin-injured hindlimb muscle of old mice by reducing SMAD4 levels. Since the miR-431 seed sequence is conserved in human SMAD4 3'UTR, miR-431 regulates the myogenic capacity of human skeletal myoblasts in the same manner. Our results suggest that age-associated miR-431 is required for the maintenance of the myogenic capability in myoblasts, thus underscoring its potential as a therapeutic target to slow down muscle aging.

• 한국응용과학기술학회지
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• 제35권2호
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• pp.509-518
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• 2018

이 연구의 목적은 신체활동이 마이오카인 발현에 미치는 영향을 보고자 문헌고찰을 하였다. 신체적인 활동은 제2형 당뇨, 심혈관질환, 대장암, 치매 및 우울증과 같은 질환을 예방하는 역할을 하고 있다. 그리고 마이오카인(myokine)은 운동 훈련에 의해 분비되는 호르몬으로 뇌성장이나 알츠하이머 같은 질환 예방에 도움을 준다. 운동수행과정에서 수축하는 근육으로부터 분비되는 항염증 마이오카인의 생성과 대사 조절에 필요한 분비 활성화가 건강증진에 중요한 요인으로 보고 있다. 인체 골격근에서 분비되는 마이오카인 가운데 IL-4, IL-6, IL-7, IL-8, IL-15 등은 근육비대(hypertrophy)와 세포(myogenesis) 및 혈관생성(angiogenesis) 등의 조절에 관여한다. IL-6는 AMPK 활성화로 인한 대사중 지방 산화를 촉진시키는 작용을 하고, IL-1Ra, IL-10 과 sTNF-R 는 염증성 싸이토카인 $TNF-{\alpha}$의 분비를 억제한다. IL-15는 저항 운동시 근수축을 통한 발현량이 증가하어 근육 성장의 중요 합성요인으로 작용한다. 한편 IL-7 및 IL-8도 신호 전달 수용체 C-X-C를 통해 혈관신생을 촉진시킨다.