The Myotube Analyzer: how to assess myogenic features in muscle stem cells

IF 5.3 2区 医学 Q2 CELL BIOLOGY Skeletal Muscle Pub Date : 2022-06-10 DOI:10.1186/s13395-022-00297-6
Noë, Simon, Corvelyn, Marlies, Willems, Sarah, Costamagna, Domiziana, Aerts, Jean-Marie, Van Campenhout, Anja, Desloovere, Kaat
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引用次数: 3

Abstract

The analysis of in vitro cultures of human adult muscle stem cells obtained from biopsies delineates the potential of skeletal muscles and may help to understand altered muscle morphology in patients. In these analyses, the fusion index is a commonly used quantitative metric to assess the myogenic potency of the muscle stem cells. Since the fusion index only partly describes myogenic potency, we developed the Myotube Analyzer tool, which combines the definition of the fusion index with extra features of myonuclei and myotubes obtained from satellite cell cultures. The software contains image adjustment and mask editing functions for preprocessing and semi-automatic segmentation, while other functions can be used to determine the features of nuclei and myotubes. The fusion index and a set of five novel parameters were tested for reliability and validity in a comparison between satellite cell cultures from children with cerebral palsy and typically developing children. These novel parameters quantified extra nucleus and myotube properties and can be used to describe nucleus clustering and myotube shape. Two analyzers who were trained in cell culture defined all parameters using the Myotube Analyzer app. Out of the six parameters, five had good reliability reflected by good intra-class correlation coefficients (> 0.75). Children with cerebral palsy were significantly different from the typically developing children (p < 0.05) for five parameters, and for three of the six parameters, these differences exceeded the minimal detectable differences. The Myotube Analyzer can be used for the analysis of fixed differentiated myoblast cultures with nuclear and MyHC staining. The app can calculate the fusion index, an already existing parameter, but also provides multiple new parameters to comprehensively describe myogenic potential in its output. The raw data used to determine these parameters are also available in the output. The parameters calculated by the tool can be used to detect differences between cultures from children with cerebral palsy and typically developing children. Since the program is open source, users can customize it to fit their own analysis requirements.
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肌管分析仪:如何评估肌肉干细胞的肌源性特征
从活组织检查中获得的成人肌肉干细胞体外培养物的分析描绘了骨骼肌的潜力,并可能有助于理解患者肌肉形态的改变。在这些分析中,融合指数是一种常用的定量指标来评估肌肉干细胞的成肌能力。由于融合指数只能部分描述肌源性,我们开发了肌管分析仪工具,该工具将融合指数的定义与从卫星细胞培养中获得的肌核和肌管的额外特征结合起来。该软件包含图像调整和掩模编辑功能,用于预处理和半自动分割,其他功能可用于确定细胞核和肌管的特征。在脑瘫儿童和正常发育儿童卫星细胞培养的比较中,对融合指数和一组五个新参数进行了可靠性和有效性测试。这些新参数量化了额外核和肌管的特性,并可用于描述核聚类和肌管形状。两名接受过细胞培养训练的分析仪使用Myotube Analyzer应用程序定义了所有参数。在六个参数中,五个具有良好的可靠性,反映了良好的类内相关系数(> 0.75)。脑瘫患儿与正常发育儿童在5个参数上存在显著差异(p < 0.05),其中3个参数的差异超过了可检测的最小差异。肌管分析仪可用于核染色和MyHC染色的固定分化成肌细胞培养物的分析。该应用程序可以计算融合指数,这是一个已经存在的参数,但也提供了多个新参数,以全面描述其输出的肌生成电位。用于确定这些参数的原始数据也可在输出中获得。该工具计算的参数可用于检测脑瘫儿童和正常发育儿童之间的文化差异。由于该程序是开源的,用户可以自定义它以适应自己的分析需求。
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来源期刊
Skeletal Muscle
Skeletal Muscle CELL BIOLOGY-
CiteScore
9.10
自引率
0.00%
发文量
25
审稿时长
12 weeks
期刊介绍: The only open access journal in its field, Skeletal Muscle publishes novel, cutting-edge research and technological advancements that investigate the molecular mechanisms underlying the biology of skeletal muscle. Reflecting the breadth of research in this area, the journal welcomes manuscripts about the development, metabolism, the regulation of mass and function, aging, degeneration, dystrophy and regeneration of skeletal muscle, with an emphasis on understanding adult skeletal muscle, its maintenance, and its interactions with non-muscle cell types and regulatory modulators. Main areas of interest include: -differentiation of skeletal muscle- atrophy and hypertrophy of skeletal muscle- aging of skeletal muscle- regeneration and degeneration of skeletal muscle- biology of satellite and satellite-like cells- dystrophic degeneration of skeletal muscle- energy and glucose homeostasis in skeletal muscle- non-dystrophic genetic diseases of skeletal muscle, such as Spinal Muscular Atrophy and myopathies- maintenance of neuromuscular junctions- roles of ryanodine receptors and calcium signaling in skeletal muscle- roles of nuclear receptors in skeletal muscle- roles of GPCRs and GPCR signaling in skeletal muscle- other relevant aspects of skeletal muscle biology. In addition, articles on translational clinical studies that address molecular and cellular mechanisms of skeletal muscle will be published. Case reports are also encouraged for submission. Skeletal Muscle reflects the breadth of research on skeletal muscle and bridges gaps between diverse areas of science for example cardiac cell biology and neurobiology, which share common features with respect to cell differentiation, excitatory membranes, cell-cell communication, and maintenance. Suitable articles are model and mechanism-driven, and apply statistical principles where appropriate; purely descriptive studies are of lesser interest.
期刊最新文献
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