Satellite cell

卫星小区
  • 文章类型: Journal Article
    肌肉生成和脂肪生成是决定家畜肌肉生长和脂肪积累的重要过程,这最终影响了他们的肉质。Hanwoo是一个受欢迎的品种,其肉类已出口到其他国家。这项研究的目的是比较卫星细胞的成肌和脂肪形成特性,Hanwoo和越南黄牛(VYC)之间的肉质。使用相同的28个月大的Hanwoo(体重:728±45kg)和VYC(体重:285±36kg)转向(每个品种n=10)。屠宰后,从腰最长肌(LL)肌肉收集组织样品用于卫星细胞分离和测定。验尸24小时后,从左侧car体侧面收集LL肌肉用于肉质分析。在相同的体外培养条件下,Hanwoo的增殖率高于VYC(p<0.05)。Hanwoo的融合指数几乎高出3倍(42.17%),与VYC相比(14.93%;p<0.05)。肌生成的表达(成肌因子5,成肌分化1,成肌素,和生肌因子6)-和脂肪生成(过氧化物酶体增殖物激活受体γ)-调节基因,Hanwoo的甘油三酯含量较高,与VYC相比(p<0.05)。Hanwoo牛肉的肌内脂肪和总单不饱和脂肪酸含量高于VYC牛肉(p<0.05)。同时,VYC肉具有较高的CIEa*和总的多不饱和脂肪酸含量(p<0.05)。总的来说,卫星细胞的体外培养特性和基因表达存在显著差异,和肉质介于Hanwoo和VYC之间。
    Myogenesis and adipogenesis are the important processes determining the muscle growth and fat accumulation livestock, which ultimately affecting their meat quality. Hanwoo is a popular breed and its meat has been exported to other countries. The objective of this study was to compare the myogenesis and adipogenesis properties in satellite cells, and meat quality between Hanwoo and Vietnamese yellow cattle (VYC). Same 28-months old Hanwoo (body weight: 728±45 kg) and VYC (body weight: 285±36 kg) steers (n=10 per breed) were used. Immediately after slaughter, tissue samples were collected from longissimus lumborum (LL) muscles for satellite cells isolation and assays. After 24 h post-mortem, LL muscles from left carcass sides were collected for meat quality analysis. Under the same in vitro culture condition, the proliferation rate was higher in Hanwoo compared to VYC (p<0.05). Fusion index was almost 3 times greater in Hanwoo (42.17%), compared with VYC (14.93%; p<0.05). The expressions of myogenesis (myogenic factor 5, myogenic differentiation 1, myogenin, and myogenic factor 6)- and adipogenesis (peroxisome proliferator-activated receptor gamma)-regulating genes, and triglyceride content were higher in Hanwoo, compared with VYC (p<0.05). Hanwoo beef had a higher intramuscular fat and total monounsaturated fatty acids contents than VYC beef (p<0.05). Whilst, VYC meat had a higher CIE a* and total polyunsaturated fatty acids content (p<0.05). Overall, there was a significant difference in the in vitro culture characteristics and genes expression of satellite cells, and meat quality between the Hanwoo and VYC.
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  • 文章类型: Observational Study
    背景:臂丛神经损伤(BPI)是导致严重功能障碍和残疾的周围神经损伤之一。如果没有及时治疗,长时间去神经支配会导致严重的肌肉萎缩。MyoD,由卫星细胞表达,是与损伤后肌肉再生过程相关的参数之一,并且假定确定神经化程序后的临床结果。本研究旨在了解手术时间(TTS)与成人臂丛神经损伤患者肱二头肌中卫星细胞MyoD表达的相关性。
    方法:在Soetomo总医院进行了横断面设计的分析性观察研究。纳入2013年5月至2015年12月期间接受手术的所有BPI患者。进行肌肉活检并使用免疫组织化学对MyoD表达进行染色。采用Pearson相关检验评估MyoD表达与TTS及年龄的相关性。
    结果:检查了22个肱二头肌样本。大多数患者是男性(81.8%),平均年龄为25.5岁。发现MyoD表达在4个月的TTS时最高,然后从9至36个月显著下降(并且稳定)。MyoD表达与TTS显著相关(r=-0.895;p=0.00),但与年龄无关(r=-0.294;p=0.184)。
    结论:我们的研究发现,从细胞的角度来看,BPI的治疗需要在MyoD表达表明的再生潜能下降之前尽早进行。
    BACKGROUND: Brachial Plexus Injury (BPI) is one of the peripheral nerve injuries which causes severe functional impairment and disability. Without prompt treatment, prolonged denervation will cause severe muscle atrophy. MyoD, which is expressed by satellite cells, is one of the parameters that relate to the regeneration process in post-injury muscle and it is presumed to determine the clinical outcome following neurotization procedure. This study aims to understand the correlation between time to surgery (TTS) and MyoD expression in satellite cells in the biceps muscle of adult brachial plexus injury patients.
    METHODS: Analytic observational study with a cross-sectional design was conducted at Dr. Soetomo General Hospital. All patients with BPI who underwent surgery between May 2013 and December 2015 were included. Muscle biopsy was taken and stained using immunohistochemistry for MyoD expression. Pearson correlation test was used to assess the correlation between MyoD expression with TTS and with age.
    RESULTS: Twenty-two biceps muscle samples were examined. Most patients are males (81.8%) with an average age of 25.5 years. MyoD expression was found to be highest at TTS of 4 months and then dropped significantly (and plateau) from 9 to 36 months. MyoD expression is significantly correlated with TTS (r=-0.895; p = 0.00) but not with age (r=-0.294; p = 0.184).
    CONCLUSIONS: Our study found, from the cellular point of view, that treatment of BPI needs to be done as early as possible before the regenerative potential - as indicated by MyoD expression - declined.
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  • 文章类型: Journal Article
    这项研究调查了通过培养从鸡和牛分离的卫星细胞获得的培养肌肉组织(CMT)的氨基酸和核苷酸相关化合物组成和味道特征,并将其与传统肉(TM)进行了比较。除缬氨酸和酪氨酸外,其他氨基酸的含量在CMT和TM之间差异显着(p<0.05)。CMT和TM在牛的谷氨酸含量之间没有显着差异,但鸡CMT中的谷氨酸含量低于TM(p<0.05)。在与核苷酸相关的化合物中,只有肌苷-5'-单磷酸(IMP)的含量是显著的,来自鸡和牛的CMT中IMP的量明显低于TM(p<0.05)。通过电子舌系统评估的味道特征存在显着差异,和鲜味,苦涩,鸡和牛的CMT和酸味值均显着低于TM(p<0.05)。本研究的结果表明,有必要开发一种卫星细胞培养方法,该方法可以增加CMT的鲜味和苦味强度,并调整生长培养基的组成,以生产具有类似于TM味道的培养肉。
    This study investigated the amino acid and nucleotide-related compound composition and taste characteristics of cultured muscle tissue (CMT) obtained by culturing satellite cells isolated from chicken and cattle and compared them to those of traditional meat (TM). The content of all amino acids except valine and tyrosine was significantly different between CMT and TM (p<0.05). The amount of glutamic acid was not significantly different between CMT and TM in cattle, but the glutamic acid in chicken CMT was lower than that of TM (p<0.05). Among the nucleotide-related compounds, only the content of inosine-5\'-monophosphate (IMP) was significant, and the amount of IMP in CMT derived from chicken and cattle was significantly lower than that of TM (p<0.05). There were significant differences in the taste characteristics assessed by an electronic tongue system, and the umami, bitterness, and sourness values of CMT were significantly lower than those of TM from both chicken and cattle (p<0.05). The results of the present study suggest that it is necessary to develop a satellite cell culture method that could increase the umami and bitterness intensity of CMT and adjust the composition of the growth medium to produce cultured meat with a taste similar to that of TM.
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  • 文章类型: Journal Article
    内源性骨骼肌发育,再生,病理学是极其复杂的过程,受局部和系统性因素的影响。了解这些机制如何发挥作用对于基础生物学和开发遗传性疾病的治疗干预措施至关重要,但也包括肌肉减少症和体积肌肉损失。离体骨骼肌模型的范围包括单独或共培养的卫星干细胞衍生的成肌细胞的二维和三维原代培养物。单个肌肉肌纤维,成肌束,和整个组织。一起,这些系统提供了获得干细胞行为机制见解的机会,细胞-细胞相互作用,在简化的系统中成熟的肌肉功能,没有混淆变量。这里,我们强调使用体外原代细胞和离体骨骼肌模型的最新进展(在过去5年中发表),总结新的见解,工具,数据集,和他们提供的筛选方法。最后,我们强调了骨骼肌知识指数级进步的机会,具有时空分辨率,通过计算机建模指导肌肉生物学和生理学的研究,并实施高含量的细胞生物学系统和离体生理学平台。
    Endogenous skeletal muscle development, regeneration, and pathology are extremely complex processes, influenced by local and systemic factors. Unpinning how these mechanisms function is crucial for fundamental biology and to develop therapeutic interventions for genetic disorders, but also conditions like sarcopenia and volumetric muscle loss. Ex vivo skeletal muscle models range from two- and three-dimensional primary cultures of satellite stem cell-derived myoblasts grown alone or in co-culture, to single muscle myofibers, myobundles, and whole tissues. Together, these systems provide the opportunity to gain mechanistic insights of stem cell behavior, cell-cell interactions, and mature muscle function in simplified systems, without confounding variables. Here, we highlight recent advances (published in the last 5 years) using in vitro primary cells and ex vivo skeletal muscle models, and summarize the new insights, tools, datasets, and screening methods they have provided. Finally, we highlight the opportunity for exponential advance of skeletal muscle knowledge, with spatiotemporal resolution, that is offered by guiding the study of muscle biology and physiology with in silico modelling and implementing high-content cell biology systems and ex vivo physiology platforms.
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  • 文章类型: Journal Article
    OBJECTIVE: Streptozotocin (STZ)-induced diabetic animal models have been widely used to study diabetic myopathy; however, non-specific cytotoxic effects of high-dose STZ have been discussed. The purpose of this study was to compare diabetic myopathy in a high-STZ model with another well-established STZ model with reduced cytotoxicity (high-fat diet (HFD) and low-dose STZ) and to identify mechanistic insights underlying diabetic myopathy in STZ models that can mimic perturbations observed in human patients with diabetic myopathy.
    METHODS: Male C57BL6 mice were injected with a single high dose of STZ (180 mg/kg, High-STZ) or were given HFD plus low-dose STZ injection (STZ, 55 mg/kg/day, five consecutive days, HFD/STZ). We characterized diabetic myopathy by histological and immunochemical analyses and conducted gene expression analysis.
    RESULTS: The high-STZ model showed a significant reduction in tibialis anterior myofiber size along with decreased satellite cell content and downregulation of inflammation response and collagen gene expression. Interestingly, blood corticosteroid levels were significantly increased in the high-STZ model, which was possibly related to lowered inflammation response-related gene expression. Further analyses using the HFD/STZ model showed downregulation of gene expression related to mitochondrial functions accompanied by a significant decrease in ATP levels in the muscles.
    CONCLUSIONS: The high-STZ model is suitable for studies regarding not only severe diabetic myopathy with excessive blood glucose but also negative impact of glucocorticoids on skeletal muscles. In contrast, the HFD/STZ model is characterized by higher immune responses and lower ATP production, which also reflects the pathologies observed in human diabetic patients.
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  • 文章类型: Journal Article
    Skeletal muscle possesses remarkable plasticity that permits functional adaptations to a wide range of signals such as motor input, exercise, and disease. Small animal models have been pivotal in elucidating the molecular mechanisms regulating skeletal muscle adaptation and plasticity. However, these small animal models fail to accurately model human muscle disease resulting in poor clinical success of therapies. Here, we review the potential of in vitro three-dimensional tissue-engineered skeletal muscle models to study muscle function, plasticity, and disease. First, we discuss the generation and function of in vitro skeletal muscle models. We then discuss the genetic, neural, and hormonal factors regulating skeletal muscle fiber-type in vivo and the ability of current in vitro models to study muscle fiber-type regulation. We also evaluate the potential of these systems to be utilized in a patient-specific manner to accurately model and gain novel insights into diseases such as Duchenne muscular dystrophy (DMD) and volumetric muscle loss. We conclude with a discussion on future developments required for tissue-engineered skeletal muscle models to become more mature, biomimetic, and widely utilized for studying muscle physiology, disease, and clinical use.
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  • 文章类型: Journal Article
    Purpose/Aim: Skeletal muscle tissue explants have been cultured and studied for nearly 100 years. These cultures, which retain complex tissue structure in an environment suited to precision manipulation and measurement, have led to seminal discoveries of the extrinsic and intrinsic mechanisms regulating contractility, metabolism and regeneration. This review discusses the two primary models of muscle explant: isolated myofiber and intact muscle.Materials and Methods: Relevant literature was reviewed and synthesized with a focus on the unique challenges and capabilities of each explant model.Results: Impactful past, current and future novel applications are discussed.Conclusions: Experiments using skeletal muscle explants have been integral to our understanding of the fundamentals of muscle physiology. As they are refined and adapted, they are poised to continue to inform the field for years to come.
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  • 文章类型: Journal Article
    卫星细胞是在出生后不久负责肌肉生长和成年期肌肉修复/再生的生肌细胞。基于卫星细胞的疗法有望治疗肌肉功能障碍。由于卫星细胞的丰度低,研究卫星细胞在技术上具有挑战性,小尺寸和解剖分散的位置之间的基底层和肌纤维的肌膜。在这篇文章中,我们提出了三种改进的方案策略,用于研究小鼠在肌肉再生的不同阶段的卫星细胞的特性:(1)新分离的单个肌纤维的免疫染色,以促进静止卫星细胞的研究,(2)在Matrigel®包被的培养皿上培养单肌纤维,以研究由卫星细胞活化启动的成肌程序,(3)在漂浮条件下培养单个肌纤维,以分析激活的卫星细胞或卫星细胞在肌纤维中的倍增时间。简而言之,与以前发表的协议相比,本文提出了一种改进的方案,该方案需要更短的实验时间和更少的费力方法,以获得更高的完整单肌纤维用于下游分析。
    Satellite cells are myogenic cells responsible for muscle growth shortly after birth and muscle repair/regeneration during adulthood. Therapies based on satellite cells hold promise for treating muscular dysfunctions. Studying satellite cells is technically challenging owing to their low abundance, small size and anatomical dispersed location between the basal lamina and the sarcolemma of myofibers. In this article, we present three improved protocol strategies for studying the properties of satellite cells of the mouse during the different stages of muscle regeneration: (1) immunostaining of freshly isolated single myofibers to facilitate the study of quiescent satellite cells, (2) cultivation of single myofibers on Matrigel®-coated dish to study the myogenesis programs initiated by satellite cell activation, and (3) cultivation of single myofibers in floating conditions to analyze activated satellite cells or the doubling time of satellite cells in myofibers. In brief, when compared to previously published protocols, this article presented an improved protocol that requires shorter experimental time and less laborious approach for higher yield of intact single myofibers for downstream analyses.
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  • 文章类型: Journal Article
    OBJECTIVE: The objective of the present study was to determine whether a denervated muscle extract (DmEx) could stimulate satellite cell response in denervated muscle.
    METHODS: Wistar rats were divided into 4 groups: normal rats, normal rats treated with DmEx, denervated rats, and denervated rats treated with DmEx. The soleus muscles were examined using immunohistochemical techniques for proliferating cell nuclear antigen, desmin, and myogenic differentiation antigen (MyoD), and electron microscopy was used for analysis of the satellite cells.
    RESULTS: The results indicate that while denervation causes activation of satellite cells, DmEx also induces myogenic differentiation of cells localized in the interstitial space and the formation of new muscle fibers. Although DmEx had a similar effect in nature on innervated and denervated muscles, this response was of greater magnitude in denervated vs. intact muscles.
    CONCLUSIONS: Our study shows that treatment of denervated rats with DmEx potentiates the myogenic response in atrophic denervated muscles.
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  • 文章类型: Journal Article
    Tibialis anterior (TA) muscle and other somite-derived limb muscles remain the prototype in skeletal muscle study. The majority of head muscles, however, develop from branchial arches and maintain a number of heterogeneities in comparison with their limb counterparts. Levator veli palatini (LVP) muscle is a deep-located head muscle responsible for breathing, swallowing and speech, and is central to cleft palate surgery, yet lacks morphological and molecular investigation. In the present study, multiscale in vivo analyses were performed to compare TA and LVP muscle in terms of their myofiber composition, in-situ stem cell population and augmentation potential. TA muscle was identified to be primarily composed of type 2B myofibers while LVP muscle primarily consisted of type 2A and 2X myofibers. In addition, LVP muscle maintained a higher percentage of centrally-nucleated myofibers and a greater population of satellite cells. Notably, TA and LVP muscle responded to exogenous Wnt7a stimulus in different ways. Three weeks after Wnt7a administration, TA muscle exhibited an increase in myofiber number and a decrease in myofiber size, while LVP muscle demonstrated no significant changes in myofiber number or myofiber size. These results suggested that LVP muscle exhibits obvious differences in comparison with TA muscle. Therefore, knowledge acquired from TA muscle studies requires further testing before being applied to LVP muscle.
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