Protocol for the isolation of mouse muscle stem cells using fluorescence-activated cell sorting.
Protocol for the isolation of mouse muscle stem cells using fluorescence-activated cell sorting.
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使用荧光激活细胞分选分离小鼠肌肉干细胞的方案。
DOI：
10.1016/j.xpro.2023.102656
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发表时间：
2023-12-15
期刊：
影响因子：
--
通讯作者：
Almada, Albert E.
中科院分区：
文献类型：
--
作者：
Elizalde, Gabriel;Munoz, Alma Zuniga;Almada, Albert E.
肌肉干细胞（Muscle Stem Cells，MuSC）是创伤后骨骼肌再生的基础。如果我们打算最大限度地发挥MuSCs的治疗潜力，我们必须进一步研究它们的分子和功能特性。在这里，我们提出了一个方案，通过两步酶促和机械解离骨骼肌结合荧光激活细胞分选（FACS）的小鼠MuSCs的分离。FACS分离的MuSC可用于各种下游应用，包括细胞培养、细胞转导、免疫荧光和基因表达测定。有关本方案使用和执行的完整详细信息，请参见Almada et al.（2021年）。H1：通过FACS分离小鼠MuSC的优化技术H2：研究MuSC命运决定的离体培养方法H3：MuSC的免疫荧光和RNA分析H4：MuSC的转导和移植方法出版商说明：进行任何实验方案都需要遵守当地机构的实验室安全和伦理指南。肌肉干细胞（Muscle Stem Cells，MuSC）是创伤后骨骼肌再生的基础。如果我们打算最大限度地发挥MuSCs的治疗潜力，我们必须进一步研究它们的分子和功能特性。在这里，我们提出了一个方案，通过两步酶促和机械解离骨骼肌结合荧光激活细胞分选（FACS）的小鼠MuSCs的分离。流式细胞术分离的MuSC可用于各种下游应用，包括细胞培养、细胞转导、免疫荧光和基因表达测定。
Muscle stem cells (MuSCs) are the building blocks for regenerating skeletal muscle after trauma. If we intend to maximize the therapeutic potential of MuSCs, we must further study their molecular and functional properties. Here, we present a protocol for the isolation of mouse MuSCs via a two-step enzymatic and mechanical dissociation of skeletal muscle coupled with fluorescence-activated cell sorting (FACS). FACS-isolated MuSCs can be used for various downstream applications including cell culture, cell transduction, immunofluorescence, and gene expression assays. For complete details on the use and execution of this protocol, please refer to Almada et al. (2021). H1: Optimized technique to isolate mouse MuSCs by FACS H2: Ex vivo culturing approaches to study MuSC fate decisions H3: Immunofluorescence and RNA analysis of MuSCs H4: Approaches for transduction and transplantation of MuSCs Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Muscle stem cells (MuSCs) are the building blocks for regenerating skeletal muscle after trauma. If we intend to maximize the therapeutic potential of MuSCs, we must further study their molecular and functional properties. Here, we present a protocol for the isolation of mouse MuSCs via a two-step enzymatic and mechanical dissociation of skeletal muscle coupled with fluorescence-activated cell sorting (FACS). FACS-isolated MuSCs can be used for various downstream applications including cell culture, cell transduction, immunofluorescence, and gene expression assays.
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