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: 1001, Q&A: 0

    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

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

    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

    Targeting the Expression of Long Noncoding RNAs in Murine Satellite Cells from Single Myofibers
    Authors:  Martina Macino, Beatrice Biferali, Andrea Cipriano, Monica Ballarino and Chiara Mozzetta, date: 11/05/2021, view: 1427, Q&A: 0

    LncRNAs have been recently implicated in the epigenetic control of muscle differentiation and their functional characterization has traditionally relied upon in vitro models of myogenic differentiation. However, the use of experimental paradigms to specifically target lncRNAs expression in muscle stem cells (MuSCs), also known as satellite cells,

    Muscle Cryoinjury and Quantification of Regenerating Myofibers in Mice
    Authors:  Yori Endo, Mehran Karvar and Indranil Sinha, date: 06/05/2021, view: 2091, Q&A: 0

    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

    Isolation, Culture, and Differentiation of Primary Myoblasts Derived from Muscle Satellite Cells
    Authors:  Kun Ho Kim, Jiamin Qiu and Shihuan Kuang, date: 07/20/2020, view: 5144, Q&A: 0
    [Abstract] The skeletal muscle is key for body mobility and motor performance, but aging and diseases often lead to progressive loss of muscle mass due to wasting or degeneration of muscle cells. Muscle satellite cells (MuSCs) represent a population of tissue stem cells residing in the skeletal muscles and are responsible for homeostatic maintenance and ...
    Analysis of Generalized Fibrosis in Mouse Tissue Sections with Masson’s Trichrome Staining
    Authors:  Diantha van de Vlekkert, Eda Machado and Alessandra d’Azzo, date: 05/20/2020, view: 4606, Q&A: 0
    [Abstract] Expansion of fibrous connective tissue and abnormal deposition of extracellular matrix (ECM) are at the basis of many fibrotic diseases. Fibrosis can occur in response to both physiological and pathological cues, including wound healing, tissue remodeling/repair and inflammation. Chronic fibrosis can lead to severe tissue damage, organ failure and ...
    A Modified Semisolid Clonal Culture for Identification of B-1 and B-2 Progenitor Colony Forming Ability of Mouse Embryonic Hemogenic Endothelial Cells
    Authors:  Michihiro Kobayashi and Momoko Yoshimoto, date: 05/05/2020, view: 2275, Q&A: 0
    [Abstract] The search for the origin of the first hematopoietic stem cells (HSCs) in the mouse embryo has been a hot topic in the field of developmental hematopoiesis. Detecting lymphoid potential is one of the supportive evidence to show the definitive hematopoietic activity of HSCs. However, the first B-lymphoid potential in the mouse embryos are reported ...
    Isolation, Purification and Characterization of Exosomes from Fibroblast Cultures of Skeletal Muscle
    Authors:  Diantha van de Vlekkert, Xiaohui Qiu, Ida Annunziata and Alessandra d'Azzo, date: 04/05/2020, view: 3979, Q&A: 0
    [Abstract] Exosomes are dynamic nanovesicles secreted by virtually all cells and are present in all biological fluids. Given their highly heterogeneous content exosomes have been implicated in many physiological and pathological processes that they exert by influencing cell-cell and cell-ECM communication. In recent years an increasing number of methods have ...
    Lipid Mixing Assay for Murine Myoblast Fusion and Other Slow Cell-cell Fusion Processes
    [Abstract] Lipid mixing (redistribution of lipid probes between fusing membranes) has been widely used to study early stages of relatively fast viral and intracellular fusion processes that take seconds to minutes. Lipid mixing assays are especially important for identification of hemifusion intermediates operationally defined as lipid mixing without content ...
    RNA Sequencing of Single Myofibers from Mus musculus
    Authors:  Darren M. Blackburn, Felicia Lazure and Vahab D. Soleimani, date: 02/20/2020, view: 4516, Q&A: 0
    [Abstract] Whole transcriptome analysis is a key method in biology that allows researchers to determine the effect a condition has on gene regulation. One difficulty in RNA sequencing of muscle is that traditional methods are performed on the whole muscle, but this captures non-myogenic cells that are part of the muscle. In order to analyze only the ...

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