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: 1037, Q&A: 0

    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

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    In situ Hybridization of miRNAs in Human Embryonic Kidney and Human Pluripotent Stem Cell-derived Kidney Organoids
    Authors:  Filipa M. Lopes, Susan J. Kimber and Ioannis Bantounas, date: 09/05/2021, view: 342, Q&A: 0

    MicroRNAs are small RNAs that negatively regulate gene expression and play an important role in fine-tuning molecular pathways during development. There is increasing interest in studying their function in the kidney, but the majority of studies to

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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: 1037, 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

    ...
    In situ Hybridization of miRNAs in Human Embryonic Kidney and Human Pluripotent Stem Cell-derived Kidney Organoids
    Authors:  Filipa M. Lopes, Susan J. Kimber and Ioannis Bantounas, date: 09/05/2021, view: 342, Q&A: 0
    [Abstract]

    MicroRNAs are small RNAs that negatively regulate gene expression and play an important role in fine-tuning molecular pathways during development. There is increasing interest in studying their function in the kidney, but the majority of studies to date use kidney cell lines and assess the total amounts of miRNAs of interest either by qPCR or by

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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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    A Simple Method to Generate Super-sensitive AID (ssAID)-based Conditional Knockouts using CRISPR-based Gene Knockout in Various Vertebrate Cell Lines
    Authors:  Kohei Nishimura and Tatsuo Fukagawa, date: 07/20/2021, view: 683, Q&A: 0
    [Abstract]

    Inducing loss of function of a target protein using methods such as gene knockout is a powerful and useful strategy for analyzing protein function in cells. In recent years, the CRISPR/Cas-9-based gene knockout technology has been widely used across a variety of eukaryotes; however, this type of simple gene knockout strategy is not applicable to

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    CRISPR/Cas9-mediated Precise SNP Editing in Human iPSC Lines
    Authors:  Hanwen Zhang and Siwei Zhang, date: 06/20/2021, view: 1451, Q&A: 0
    [Abstract]

    Human induced pluripotent stem cells (hiPSCs) have been extensively used in the fields of developmental biology and disease modeling. CRISPR/Cas9 gene editing in iPSC lines often has a low frequency, which hampers its application in precise allele editing of disease-associated single nucleotide polymorphisms (SNPs), especially those in the

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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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    Generation of Mouse Pluripotent Stem Cell-derived Trunk-like Structures: An in vitro Model of Post-implantation Embryogenesis
    [Abstract]

    Post-implantation mammalian embryogenesis involves profound molecular, cellular, and morphogenetic changes. The study of these highly dynamic processes is complicated by the limited accessibility of in utero development. In recent years, several complementary in vitro systems comprising self-organized assemblies of mouse embryonic stem cells, such

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    Generation and Maintenance of Homogeneous Human Midbrain Organoids
    Authors:  Henrik Renner, Martha Grabos, Hans R. Schöler and Jan M. Bruder, date: 06/05/2021, view: 2046, Q&A: 0
    [Abstract]

    Three-dimensional cell cultures (“organoids”) promise to better recapitulate native tissue physiology than traditional 2D cultures and are becoming increasingly interesting for disease modeling and compound screening efforts. While a number of protocols for the generation of neural organoids have been published, most protocols require

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