Stem Cell

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    Protocols in Current Issue
    Using BODIPY FL-Sphingolipid Analogs to Study Sphingolipid Metabolism in Mouse Embryonic Stem Cells
    Authors:  Wei Fan and Xiaoling Li, date: 11/20/2022, view: 347, Q&A: 0

    Sphingolipids are important structural components of cellular membranes. They also function as prominent signaling molecules to control a variety of cellular events, such as cell growth, differentiation, and apoptosis. Impaired sphingolipid

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    Using BODIPY FL-Sphingolipid Analogs to Study Sphingolipid Metabolism in Mouse Embryonic Stem Cells
    Authors:  Wei Fan and Xiaoling Li, date: 11/20/2022, view: 347, Q&A: 0
    [Abstract]

    Sphingolipids are important structural components of cellular membranes. They also function as prominent signaling molecules to control a variety of cellular events, such as cell growth, differentiation, and apoptosis. Impaired sphingolipid metabolism, particularly defects in sphingolipid degradation, has been associated with many human diseases.

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    Generation of iMyoblasts from Human Induced Pluripotent Stem Cells
    [Abstract]

    Skeletal muscle stem cells differentiated from human-induced pluripotent stem cells (hiPSCs) serve as a uniquely promising model system for investigating human myogenesis and disease pathogenesis, and for the development of gene editing and regenerative stem cell therapies. Here, we present an effective and reproducible transgene-free protocol for

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    Isolation and ex vivo Expansion of Limbal Mesenchymal Stromal Cells
    Authors:  Naresh Polisetti, Lyne Sharaf, Thomas Reinhard and Günther Schlunck, date: 07/20/2022, view: 893, Q&A: 0
    [Abstract]

    Limbal mesenchymal stromal cells (LMSC), a cellular component of the limbal stem cell niche, have the capability of determining the fate of limbal epithelial progenitor cells (LEPC), which are responsible for the homeostasis of corneal epithelium. However, the isolation of these LMSC has proven to be difficult due to the small fraction of LMSC in

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    Gene Expression Analysis in Stem Cell-derived Cortical Neuronal Cultures Using Multi-well SYBR Green Quantitative PCR Arrays
    Authors:  Vasavi Nallur Srinivasaraghavan, Faria Zafar and Birgitt Schüle, date: 07/20/2022, view: 1156, Q&A: 0
    [Abstract]

    To optimize differentiation protocols for stem cell-based in vitro modeling applications, it is essential to assess the change in gene expression during the differentiation process. This allows controlling its differentiation efficiency into the target cell types. While RNA transcriptomics provides detail at a larger scale, timing and cost are

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    Assessing the Presence of Hematopoietic Stem and Progenitor Cells in Mouse Spleen
    Authors:  Isabelle J. Marié, Lara Brambilla and David E. Levy, date: 06/05/2022, view: 1363, Q&A: 0
    [Abstract]

    Transplantation of hematopoietic material into recipient mice is an assay routinely used to determine the presence and function of hematopoietic stem and progenitor cells (HSPCs) in vivo. The principle of the method is to transplant donor cells being tested for HSPCs into a recipient mouse following bone marrow ablation and testing for

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    Click-iT® Plus OPP Alexa Fluor® Protein Synthesis Assay in Embryonic Cells
    [Abstract]

    This protocol describes a method to assess relative changes in the level of global protein synthesis in the preimplantation embryo using the Click-iT® Plus OPP Protein Synthesis Assays. In this assay, O-propargyl-puromycin (OPP), an analog of puromycin, is efficiently incorporated into the nascent polypeptide of newly translated proteins in

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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: 1490, 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: 1262, Q&A: 1
    [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: 1697, 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: 1367, 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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