Analyses of Mesenchymal Progenitors in Skeletal Muscle by Fluorescence-Activated Cell Sorting and Tissue Clearing
Analyses of Mesenchymal Progenitors in Skeletal Muscle by Fluorescence-Activated Cell Sorting and Tissue Clearing
复制标题
通过荧光激活细胞分选和组织透明化分析骨骼肌中的间充质祖细胞
DOI：
10.1007/978-1-0716-3036-5_9
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Uezumi A
中科院分区：
文献类型：
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作者：
Ikemoto-Uezumi M;Kurosawa T;Minato K;Uezumi A
间充质祖细胞是居住在骨骼肌间质中的祖细胞群体，在脂肪渗透、纤维化和异位骨化等发病机制中起重要作用。除了它们的病理作用外，间充质前体细胞也被证明在成功的肌肉再生和维持肌肉内稳方面发挥着重要作用。因此，对这些祖细胞的详细和准确的分析对于肌肉疾病和健康的研究是必不可少的。在这里，我们描述了一种基于PDGFRα表达的纯化间充质祖细胞的方法，PDGFRDNA是间充质祖细胞的特异性和公认的标志物，使用荧光激活细胞分选技术(FAC)。纯化的细胞可以用于几个下游实验，包括细胞培养、细胞移植和基因表达分析。我们还描述了利用组织透明技术对间充质祖细胞进行整体三维成像的方法。本文描述的方法为研究骨骼肌间充质祖细胞提供了一个强大的平台。
Mesenchymal progenitors, which are resident progenitor populations residing in skeletal muscle interstitial space, contribute to pathogeneses such as fat infiltration, fibrosis, and heterotopic ossification. In addition to their pathological roles, mesenchymal progenitors have also been shown to play important roles for successful muscle regeneration and homeostatic muscle maintenance. Therefore, detailed and accurate analyses of these progenitors are essential for the research on muscle diseases and health. Here, we describe a method for purification of mesenchymal progenitors based on the expression of PDGFRα, which is a specific and well-established marker for mesenchymal progenitors, using fluorescence-activated cell sorting (FACS). Purified cells can be used in several downstream experiments including cell culture, cell transplantation, and gene expression analysis. We also describe the method for whole-mount 3-dimensional imaging of mesenchymal progenitors by utilizing tissue clearing. The methods described herein provide a powerful platform for studying mesenchymal progenitors in skeletal muscle.