Stem Cell

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    Protocols in Current Issue
    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: 1038, 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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    Isolation of Single Cells from Mouse Periodontal Ligament
    Authors:  Jing Zhao and Paul Sharpe, date: 08/20/2021, view: 836, Q&A: 0
    [Abstract]

    The periodontal ligament (PDL) is an essential tissue connecting teeth and bone. It is a complex tissue specifically designed to absorb the forces of mastication; analysis of its multiple cell populations is important to understand its function and the cell changes associated with periodontal disease. Cells in the periodontal ligament are not

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    Neutral Comet Assay to Detect and Quantitate DNA Double-Strand Breaks in Hematopoietic Stem Cells
    Authors:  Irene Mariam Roy, Pon Sowbhagya Nadar and Satish Khurana, date: 08/20/2021, view: 448, Q&A: 0
    [Abstract]

    In vertebrates, hematopoietic stem cells (HSCs) regulate the supply of blood cells throughout the lifetime and help to maintain homeostasis. Due to their long lifespan, genetic integrity is paramount for these cells, and accordingly, a number of stem cell-specific mechanisms are employed. However, HSCs tend to show more DNA damage with increasing

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    Mouse Periosteal Cell Culture, in vitro Differentiation, and in vivo Transplantation in Tibial Fractures
    [Abstract]

    The periosteum covering the outer surface of bone contains skeletal stem/progenitor cells that can efficiently form cartilage and bone during bone repair. Several methods have been described to isolate periosteal cells based on bone scraping and/or enzymatic digestion. Here, we describe an explant culture method to isolate periosteum-derived

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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: 1380, 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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    Ex vivo Tissue Culture Protocols for Studying the Developing Neocortex
    Authors:  Takashi Namba, Christiane Haffner and Wieland B. Huttner, date: 05/20/2021, view: 1865, Q&A: 0
    [Abstract]

    The size of the neocortex and its morphology are highly divergent across mammalian species. Several approaches have been utilized for the analysis of neocortical development and comparison among different species. In the present protocol (Note: This protocol requires basic knowledge of brain anatomy), we describe three ex vivo neocortical

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    Generation of the Compression-induced Dedifferentiated Adipocytes (CiDAs) Using Hypertonic Medium
    [Abstract]

    Current methods to obtain mesenchymal stem cells (MSCs) involve sampling, culturing, and expanding of primary MSCs from adipose, bone marrow, and umbilical cord tissues. However, the drawbacks are the limited numbers of total cells in MSC pools, and their decaying stemness during in vitro expansion. As an alternative resource, recent ceiling

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    Establishing an Adult Mouse Brain Hippocampal Organotypic Slice Culture System that Allows for Tracing and Pharmacological Manipulation of ex vivo Neurogenesis
    Authors:  Steffen Mayerl and Charles ffrench-Constant, date: 01/05/2021, view: 2066, Q&A: 0
    [Abstract]

    The function of the hippocampus depends on the process of adult hippocampal neurogenesis which underpins the exceptional neural plasticity of this structure, and is also frequently affected in CNS pathologies. Thus, manipulation of this process represents an important therapeutic goal. To identify potential strategies, organotypic adult brain

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    Isolation of Extracellular Vesicles Derived from Mesenchymal Stromal Cells by Ultracentrifugation
    [Abstract] Extracellular vesicles (EVs) are a heterogeneous group of membranous vesicles that differ on their biogenesis and release pathways, such as exosomes, microvesicles and apoptotic bodies. They are involved in cell-to-cell communication delivering signal molecules (proteins, nucleic acids, lipids, etc.) that can regulate different ...
    Fluidigm Based Single-cell Gene Expression Library Preparation from Patient-derived Small Intestinal Organoids
    Authors:  Kohei Suzuki and Ryuichi Okamoto, date: 10/05/2020, view: 1905, Q&A: 0
    [Abstract] In this protocol, we describe our methods to isolate crypts from patients' biopsy samples and to culture human intestinal stem cells as it’s called “organoid.” Beyond that, we describe how to dissociate organoids cells into single cells for single-cell analysis as a further application. This protocol should provide investigators sufficient tools ...



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