Stem Cell

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    Protocols in Current Issue
    An in vitro Mechanical Damage Model of Isolated Myofibers in a Floating Culture Condition
    Authors:  Yoshifumi Tsuchiya and Yusuke Ono, date: 01/05/2022, view: 1133, Q&A: 0
    [Abstract]

    Muscle stem cells (satellite cells), located on the surface of myofibers, are rapidly activated from a quiescent state following skeletal muscle injury. Although satellite cell activation is an initial step in muscle regeneration, the stimulation of satellite cell activation by muscle injury remains to be elucidated. We recently established an in

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    A Simple Method for the Isolation and in vitro Expansion of Highly Pure Mouse and Human Satellite Cells
    [Abstract]

    Satellite cells (SCs) are muscle stem cells capable of regenerating injured muscle. The study of their functional potential depends on the availability of methods for the isolation and expansion of pure SCs, which retain myogenic properties after serial passages in vitro. Here, we describe a protocol for the isolation and in vitro expansion of

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    Whole-mount Staining of Mouse Diaphragm Neuromuscular Junctions
    Authors:  Rula Sha, Zhenzhen Wang, Xue You, Yuguo Liu, Zhiqin Xie and Ying Feng, date: 11/05/2021, view: 1221, Q&A: 0
    [Abstract]

    The neuromuscular junction (NMJ) is a specialized synapse that connects the terminal end of a motor neuron and a skeletal muscle fiber. Defects in NMJ cause abnormalities of neuromuscular transmission, leading to NMJ disorders. The mammalian diaphragm muscle is essential for respiration and has been widely used to study NMJ formation. Here, we

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    Isolation of Myofibres and Culture of Muscle Stem Cells from Adult Zebrafish
    Authors:  Massimo Ganassi, Peter S. Zammit and Simon M. Hughes, date: 09/05/2021, view: 3153, Q&A: 0
    [Abstract]

    Skeletal muscles generate force throughout life and require maintenance and repair to ensure efficiency. The population of resident muscle stem cells (MuSCs), termed satellite cells, dwells beneath the basal lamina of adult myofibres and contributes to both muscle growth and regeneration. Upon exposure to activating signals, MuSCs proliferate to

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    Muscle Cryoinjury and Quantification of Regenerating Myofibers in Mice
    Authors:  Yori Endo, Mehran Karvar and Indranil Sinha, date: 06/05/2021, view: 2173, Q&A: 0
    [Abstract]

    Cryoinjury, or injury due to freezing, is a method of creating reproducible, local injuries in skeletal muscle. This method allows studying the regenerative response following muscle injuries in vivo, thus enabling the evaluation of local and systemic factors that influence the processes of myofiber regeneration. Cryoinjuries are applicable to the

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    Isolation and Culture of Single Myofiber and Immunostaining of Satellite Cells from Adult C57BL/6J Mice
    Authors:  Shujie Chen, Hongrong Ding, Xiangping Yao and Liwei Xie, date: 07/20/2019, view: 6041, Q&A: 0
    [Abstract] Myofiber isolation followed with ex vivo culture could recapitulate and visualize satellite cells (SCs) activation, proliferation, and differentiation. This approach could be taken to understand the physiology of satellite cells and the molecular mechanism of regulatory factors, in terms of the involvement of intrinsic factors over SCs ...
    Microarray, IPA and GSEA Analysis in Mice Models
    [Abstract] This protocol details a method to analyze two tissue samples at the transcriptomic level using microarray analysis, ingenuity pathway analysis (IPA) and gene set enrichment analysis (GSEA). Methods such as these provide insight into the mechanisms underlying biological differences across two samples and thus can be applied to interrogate a variety ...
    A Method to Injure, Dissect and Image Indirect Flight Muscle of Drosophila
    Authors:  Kunal Chakraborty, K. VijayRaghavan and Rajesh Gunage, date: 05/20/2018, view: 7477, Q&A: 0
    [Abstract] Inducing an injury specifically to Drosophila flight muscles is a difficult task, owing to the small size of the muscles and the presence of the cuticle. The protocol described below provides an easy and reproducible method to induce injury in the Drosophila flight muscles.
    Mouse Satellite Cell Isolation and Transplantation
    [Abstract] Satellite cell (SC) transplantation represents a powerful strategy to investigate SC biology during muscle regeneration. We described here a protocol for SC isolation from green fluorescent protein (GFP)-expressing mice and their transplantation into murine muscles. This procedure was originally used to assess the effects of the hormone unacylated ...
    Isolation of Primary Human Skeletal Muscle Cells
    Authors:  Janelle M. Spinazzola and Emanuela Gussoni, date: 11/05/2017, view: 7900, Q&A: 0
    [Abstract] Primary myoblast culture is a valuable tool in research of muscle disease, pathophysiology, and pharmacology. This protocol describes techniques for dissociation of cells from human skeletal muscle biopsies and enrichment for a highly myogenic population by fluorescence-activated cell sorting (FACS). We also describe methods for assessing ...



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