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: 102, 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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    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: 1349, 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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    APEX-mediated Proximity Labeling of Proteins in Cells Targeted by Extracellular Vesicles
    Authors:  Lu Song, Jun Chen, Angela Sun and Randy Schekman, date: 11/05/2021, view: 1825, Q&A: 0
    [Abstract]

    Extracellular vesicles (EVs) are thought to mediate intercellular communication through the delivery of cargo proteins and RNA to target cells. The uptake of EVs is often followed visually using lipophilic-dyes or fluorescently-tagged proteins to label membrane constituents that are then internalized into recipient cells (Christianson et al.,

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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: 1603, 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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    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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    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: 2277, 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 ...
    Differentiation of Mouse Embryonic Stem Cells to Neuronal Cells Using Hanging Droplets and Retinoic Acid
    Authors:  Jeroen Witteveldt and Sara Macias, date: 11/05/2019, view: 4121, Q&A: 0
    [Abstract] Controlled differentiation of embryonic stem cells is an essential tool in stem cell research. In this protocol, we describe a simple differentiation protocol involving the induction of embryoid body formation in mouse embryonic stem cells (mESC) using hanging droplets, followed by differentiation into a neuronal lineage.
    An Improved Staining Method for Low Signal LacZ Reporter Detection in Mouse Embryos
    Authors:  Xiaopeng Shen, Feng Xu, Shen Wu, Meng Li, Jingyi Zhang, Rui Liang and Yu Liu, date: 03/05/2019, view: 4980, Q&A: 0
    [Abstract] In many fields of biology, especially in the field of developmental biology, LacZ reporter staining is an approach used to monitor gene expression patterns. In the LacZ reporter system, the LacZ gene is inserted in the endogenous location of the target gene via gene knock-in or by constructing a transgenic cassette in ...
    Isolation of Neural Stem Cells from the Embryonic Mouse Hippocampus for in vitro Growth or Engraftment into a Host Tissue
    Authors:  Oksana Rybachuk, Olga Kopach, Tetyana Pivneva and Vitaliy Kyryk, date: 02/20/2019, view: 6758, Q&A: 0
    [Abstract] For both stem cell research and treatment of the central nervous system disorders, neural stem/progenitor cells (NSPCs) represent an important breakthrough tool. In the expanded stem cell-based therapy use, NSPCs not only provide a powerful cell source for neural cell replacement but a useful model for developmental biology research. Despite ...
    Quantification of Mouse Hematopoietic Progenitors’ Formation Using Time-lapse Microscopy and Image Analysis
    [Abstract] In vitro differentiation of mouse embryonic stem cells (mESCs) towards blood cells constitutes a well-established system to study the endothelial-to-hematopoietic transition (EHT) at the onset of blood development. Assessing the emergence of small non-adherent round blood cells in the culture without disturbing it is essential to evaluate ...



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