Division of Life Science, Center for Stem Cell Research, HKUST-Nan Fung Life Sciences Joint Laboratory, State Key Laboratory of Molecular Neuroscience, Molecular Neuroscience Center, Hong Kong University of Science and Technology, Kowloon, Hong Kong, China.
Institut NeuroMyoGène, Université Claude Bernard Lyon 1, Lyon, France.
Methods Mol Biol. 2023;2640:369-395. doi: 10.1007/978-1-0716-3036-5_26.
Skeletal muscle possesses a remarkable regenerative capacity, mainly relying on a population of undifferentiated and unipotent muscle progenitors, called muscle stem cells (MuSCs) or satellite cells, and their interplay with various cell types within the niche. Investigating the cellular composition of skeletal muscle tissues and the heterogeneity among various cell populations is crucial to the unbiased understanding of how cellular networks work in harmony at the population level in the context of skeletal muscle homeostasis, regeneration, aging, and diseases. As opposed to probing the average profile in a cell population, single-cell RNA-seq has unlocked access to the transcriptomic landscape characterization of individual cells in a highly parallel manner. This chapter describes the workflow for single-cell transcriptomic analysis of mononuclear cells in skeletal muscle by taking advantage of the droplet-based single-cell RNA-seq platform, Chromium Single Cell 3' solution from 10x Genomics. Using this protocol, we can reveal insights into muscle-resident cell-type identities, which can be exploited to study the muscle stem cell niche further.
骨骼肌具有显著的再生能力，主要依赖于一群未分化的、单能的肌肉祖细胞，称为肌肉干细胞（MuSCs）或卫星细胞，以及它们与龛内各种细胞类型的相互作用。研究骨骼肌组织的细胞组成以及各种细胞群体之间的异质性，对于在骨骼肌稳态、再生、衰老和疾病的背景下，从群体水平上了解细胞网络如何协调工作至关重要。单细胞 RNA 测序的出现与应用，与探测细胞群体的平均特征不同，它为我们提供了一种高通量的方法，可以对单个细胞的转录组景观进行特征描述。本章介绍了一种利用基于液滴的单细胞 RNA 测序平台（10x Genomics 的 Chromium Single Cell 3' solution），对骨骼肌单核细胞进行单细胞转录组分析的工作流程。通过使用该方案，我们可以深入了解肌肉驻留细胞类型的特征，这可以进一步用于研究肌肉干细胞龛。
