Stem Cell

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    Protocols in Current Issue
    Microarray, IPA and GSEA Analysis in Mice Models
    [Abstract] This protocol details a method to analyze two tissue samples at the transcriptomic level using microarray analysis, ingenuity pathway analysis (IPA) and gene set enrichment analysis (GSEA). Methods such as these provide insight into the mechanisms underlying biological differences across two samples and thus can be applied to interrogate a variety ...
    A Method to Injure, Dissect and Image Indirect Flight Muscle of Drosophila
    Authors:  Kunal Chakraborty, K. VijayRaghavan and Rajesh Gunage, date: 05/20/2018, view: 2566, Q&A: 0
    [Abstract] Inducing an injury specifically to Drosophila flight muscles is a difficult task, owing to the small size of the muscles and the presence of the cuticle. The protocol described below provides an easy and reproducible method to induce injury in the Drosophila flight muscles.
    Mouse Satellite Cell Isolation and Transplantation
    [Abstract] Satellite cell (SC) transplantation represents a powerful strategy to investigate SC biology during muscle regeneration. We described here a protocol for SC isolation from green fluorescent protein (GFP)-expressing mice and their transplantation into murine muscles. This procedure was originally used to assess the effects of the hormone unacylated ...
    Isolation of Primary Human Skeletal Muscle Cells
    Authors:  Janelle M. Spinazzola and Emanuela Gussoni, date: 11/05/2017, view: 3367, Q&A: 0
    [Abstract] Primary myoblast culture is a valuable tool in research of muscle disease, pathophysiology, and pharmacology. This protocol describes techniques for dissociation of cells from human skeletal muscle biopsies and enrichment for a highly myogenic population by fluorescence-activated cell sorting (FACS). We also describe methods for assessing ...
    Isolation, Culturing, and Differentiation of Primary Myoblasts from Skeletal Muscle of Adult Mice
    Authors:  Lubna Hindi, Joseph D. McMillan, Dil Afroze, Sajedah M. Hindi and Ashok Kumar, date: 05/05/2017, view: 11879, Q&A: 4
    [Abstract] Myogenesis is a multi-step process that leads to the formation of skeletal muscle during embryonic development and repair of injured myofibers. In this process, myoblasts are the main effector cell type which fuse with each other or to injured myofibers leading to the formation of new myofibers or regeneration of skeletal muscle in adults. Many ...
    Quantitative 3D Time Lapse Imaging of Muscle Progenitors in Skeletal Muscle of Live Mice
    Authors:  Micah T. Webster, Tyler Harvey and Chen-Ming Fan, date: 12/20/2016, view: 5025, Q&A: 0
    [Abstract] For non-optically clear mammalian tissues, it is now possible to use multi-photon microscopy to penetrate deep into the tissue and obtain detailed single cell images in a live animal, i.e., intravital imaging. This technique is in principle applicable to any fluorescently marked cell, and we have employed it to observe stem cells during ...
    Isolation, Culture, and Staining of Single Myofibers
    Authors:  Yann Simon Gallot, Sajedah M. Hindi, Aman K. Mann and Ashok Kumar, date: 10/05/2016, view: 6984, Q&A: 0
    [Abstract] Adult skeletal muscle regeneration is orchestrated by a specialized population of adult stem cells called satellite cells, which are localized between the basal lamina and the plasma membrane of myofibers. The process of satellite cell-activation, proliferation, and subsequent differentiation that occurs during muscle regeneration can be ...
    FACS-based Satellite Cell Isolation From Mouse Hind Limb Muscles
    Authors:  Anastasia Gromova, Matthew T Tierney and Alessandra Sacco, date: 08/20/2015, view: 7323, Q&A: 0
    [Abstract] Fluorescence Activated Cell Sorting (FACS) is a sensitive and accurate method for purifying satellite cells, or muscle stem cells, from adult mouse skeletal muscle (Liu et al., 2013; Sacco et al., 2008; Tierney et al., 2014). Mechanical and enzymatic digestion of hind limb muscles releases mononuclear muscle cells into ...
    Isolation of FAP Cells from Mouse Dystrophic Skeletal Muscle Using Fluorescence Activated Cell Sorting
    Authors:  Nicoletta Cordani, Viviana Pisa, Laura Pozzi and Clara Sciorati, date: 11/20/2014, view: 8038, Q&A: 1
    [Abstract] A population of muscle resident CD45-, CD31- cells expressing the mesenchymal PDGF receptor alpha (PDGFRα) as well as Sca-1 was first isolated in healthy mouse muscles in Uezumi et al. (2010). In the same year, Joe et al. (2010) identified and purified fibro-adipogenic precursors (FAPs), cells located into ...
    Preparation of Adult Mouse Muscle Stem Cells
    Author:  Ju Li, date: 12/20/2013, view: 7017, Q&A: 0
    [Abstract] Muscle stem cells are adult stem cells responsible for muscle development, growth and regeneration. Current knowledge suggests those cells are heterogeneous population shared their position between the sarcolemma and basal lamina of muscle fibers. This protocol describes the technique to dissociate and collect the stem cells from skeletal muscle ...



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