Stem Cell

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    Protocols in Current Issue
    Optimized CRISPR-Cas9-based Strategy for Complex Gene Targeting in Murine Embryonic Stem Cells for Germline Transmission
    Authors:  Thomas J. O'Neill, Daniel Krappmann and Andreas Gewies, date: 05/20/2022, view: 70, Q&A: 0

    Although CRISPR-Cas9 genome editing can be performed directly in single-cell mouse zygotes, the targeting efficiency for more complex modifications such as the insertion of two loxP sites, multiple mutations in cis, or the precise insertion or

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    Optimized CRISPR-Cas9-based Strategy for Complex Gene Targeting in Murine Embryonic Stem Cells for Germline Transmission
    Authors:  Thomas J. O'Neill, Daniel Krappmann and Andreas Gewies, date: 05/20/2022, view: 70, Q&A: 0
    [Abstract]

    Although CRISPR-Cas9 genome editing can be performed directly in single-cell mouse zygotes, the targeting efficiency for more complex modifications such as the insertion of two loxP sites, multiple mutations in cis, or the precise insertion or deletion of longer DNA sequences often remains low (Cohen, 2016). Thus, targeting and validation of

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    Isolation and Culture of Cranial Neural Crest Cells from the First Branchial Arch of Mice
    Authors:  Hiroki Ueharu, Jingwen Yang, Yoshihiro Komatsu and Yuji Mishina, date: 04/05/2022, view: 456, Q&A: 0
    [Abstract]

    Craniofacial anomalies (CFA) are a diverse group of deformities, which affect the growth of the head and face. Dysregulation of cranial neural crest cell (NCC) migration, proliferation, differentiation, and/or cell fate specification have been reported to contribute to CFA. Understanding of the mechanisms through which cranial NCCs contribute

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    Differentiation of Human Induced Pluripotent Stem Cell into Macrophages
    Authors:  Harriet Douthwaite, Aitor Bermejo Arteagabeitia and Subhankar Mukhopadhyay, date: 03/20/2022, view: 836, Q&A: 0
    [Abstract]

    As a model to interrogate human macrophage biology, macrophages differentiated from human induced pluripotent stem cells (hiPSCs) transcend other existing models by circumventing the variability seen in human monocyte-derived macrophages, whilst epitomizing macrophage phenotypic and functional characteristics over those offered by macrophage-like

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    Skeletal Stem Cell Isolation from Cranial Suture Mesenchyme and Maintenance of Stemness in Culture
    Authors:  Takamitsu Maruyama, Hsiao-Man Ivy Yu and Wei Hsu, date: 03/05/2022, view: 578, Q&A: 0
    [Abstract]

    Skeletal stem cells residing in the suture mesenchyme are responsible for calvarial development, homeostatic maintenance, and injury-induced repair. These naïve cells exhibit long-term self-renewal, clonal expansion, and multipotency. They possess osteogenic abilities to regenerate bones in a cell-autonomous manner and can directly replace

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    Efficient Method to Differentiate Mouse Embryonic Stem Cells into Macrophages in vitro
    Authors:  Qimin Hai, Juying Han, Sophia Wells and Jonathan D. Smith, date: 02/05/2022, view: 1345, Q&A: 0
    [Abstract]

    Macrophages are key cells in the innate immune system and play a role in a variety of diseases. However, macrophages are terminally differentiated and difficult to manipulate genetically via transfection or through CRISPR-Cas9 gene editing. To overcome this limitation, we provide a simplified protocol for the generation of mouse embryonic stem

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    Flow Cytometry Analysis of Planarian Stem Cells Using DNA and Mitochondrial Dyes
    Authors:  Mohamed Mohamed Haroon, Praveen Kumar Vemula and Dasaradhi Palakodeti, date: 01/20/2022, view: 931, Q&A: 0
    [Abstract]

    Planarians are free-living flatworms that emerged as a crucial model system to understand regeneration and stem cell biology. The ability to purify neoblasts, the adult stem cell population of planaria, through fluorescence-activated cell sorting (FACS) has tremendously increased our understanding of pluripotency, specialization, and

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    From 3D to 2D: Harmonization of Protocols for Two-dimensional Cultures on Cell Culture Inserts of Intestinal Organoids from Various Species
    [Abstract]

    In the expanding field of intestinal organoid research, various protocols for three- and two-dimensional organoid-derived cell cultures exist. Two-dimensional organoid-derived monolayers are used to overcome some limitations of three-dimensional organoid cultures. They are increasingly used also in infection research, to study physiological

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    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
    [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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    Live-cell Imaging and Analysis of Germline Stem Cell Mitosis in Caenorhabditis elegans
    Authors:  Réda M. Zellag, Yifan Zhao and Abigail R. Gerhold, date: 01/05/2022, view: 1241, Q&A: 0
    [Abstract]

    Model organisms offer the opportunity to decipher the dynamic and complex behavior of stem cells in their native environment; however, imaging stem cells in situ remains technically challenging. C. elegans germline stem cells (GSCs) are distinctly accessible for in situ live imaging but relatively few studies have taken advantage of this

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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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