Stem Cell

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    Protocols in Current Issue
    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: 332, Q&A: 0

    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

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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: 332, 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: 1183, 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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    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: 1807, 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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    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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    Muscle Cryoinjury and Quantification of Regenerating Myofibers in Mice
    Authors:  Yori Endo, Mehran Karvar and Indranil Sinha, date: 06/05/2021, view: 1230, 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: 1692, 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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    Single or Repeated Ablation of Mouse Olfactory Epithelium by Methimazole
    Authors:  Sofia Håglin, Staffan Bohm and Anna Berghard, date: 04/20/2021, view: 1892, Q&A: 0
    [Abstract]

    Odor-detecting olfactory sensory neurons residing in the nasal olfactory epithelium (OE) are the only neurons in direct contact with the external environment. As a result, these neurons are subjected to chemical, physical, and infectious insults, which may be the underlying reason why neurogenesis occurs in the OE of adult mammals. This feature

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    Generation of Human iPSC-derived Neural Progenitor Cells (NPCs) as Drug Discovery Model for Neurological and Mitochondrial Disorders
    Authors:  Annika Zink, Pawel Lisowski and Alessandro Prigione, date: 03/05/2021, view: 2261, Q&A: 1
    [Abstract]

    The high attrition rate in drug development processes calls for additional human-based model systems. However, in the context of brain disorders, sampling live neuronal cells for compound testing is not applicable. The use of human induced pluripotent stem cells (iPSCs) has revolutionized the field of neuronal disease modeling and drug discovery.

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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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    Rapid and Simplified Induction of Neural Stem/Progenitor Cells (NSCs/NPCs) and Neurons from Human Induced Pluripotent Stem Cells (hiPSCs)
    Authors:  Ryutaro Kajihara, Tadahiro Numakawa and Takumi Era, date: 02/05/2021, view: 1878, Q&A: 0
    [Abstract]

    Human induced pluripotent stem cells (iPSCs) and their progeny displaying tissue-specific characteristics have paved the way for regenerative medicine and research in various fields such as the elucidation of the pathological mechanism of diseases and the discovery of drug candidates. iPSC-derived neurons are particularly valuable as it is

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