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    Protocols in Current Issue
    Alphavirus Purification Using Low-speed Spin Centrifugation
    [Abstract] Chemical and sedimentation procedures are used to purify virus particles. While these approaches are successful for wild-type viruses, they are often not feasible for purifying mutant viruses with assembly defects. We combined two published methods (Atasheva et al., 2013; Moller-Tank et al., 2013), to generate a protocol that ...
    Isolation and Establishment of Mesenchymal Stem Cells from Wharton’s Jelly of Human Umbilical Cord
    Authors:  Umesh Goyal, Chitra Jaiswal and Malancha Ta, date: 02/20/2018, view: 763, Q&A: 3
    [Abstract] Mesenchymal stem cells (MSCs) are currently considered as ‘medicinal signaling cells’ and a promising resource in regard to cell-based regenerative therapy. Umbilical cord is a human term perinatal tissue which is easily attainable, and a promising source of stem cells with no associated ethical concerns. MSCs have been isolated from different ...
    Culture and Nucleofection of Postnatal Day 7 Cortical and Cerebellar Mouse Astroglial Cells
    Authors:  Malek Chouchane and Marcos Romualdo Costa, date: 02/05/2018, view: 515, Q&A: 0
    [Abstract] Lineage reprogramming of astroglial cells isolated from different brain regions leads to the generation of different neuronal subtypes. This protocol describes the isolation and culture of neocortical and cerebellar astrocytes from postnatal mice. We also present a comprehensive description of the main steps towards successful gene delivery in ...
    Generation of Chemically Induced Liver Progenitors (CLiPs) from Rat Adult Hepatocytes
    Authors:  Takeshi Katsuda, Kazunori Hosaka and Takahiro Ochiya, date: 01/20/2018, view: 835, Q&A: 0
    [Abstract] Primary mature hepatocytes (MHs) or their progenitor cells are candidate cell sources for cell transplantation therapy in severe liver diseases. However, stable culture of these cells or generation of equivalent cells from pluripotent stem cells has been limited. Using a cocktail of small molecules that we previously found useful in stable culture ...
    Isolation and Purification of Viruses Infecting Cyanobacteria Using a Liquid Bioassay Approach
    Authors:  Caroline Chénard and Amy M Chan, date: 01/20/2018, view: 710, Q&A: 0
    [Abstract] The following protocol describes the isolation and purification of viruses infecting cyanobacteria using a liquid bioassay approach. Viruses infecting cyanobacteria are also known as cyanophages. This protocol was written specifically for the isolation of cyanophages infecting freshwater cyanobacteria particularly, cyanobacteria that cannot be ...
    Human Endometrial Stem Cell Isolation from Endometrium and Menstrual Blood
    Authors:  Shima Tavakol, Freshteh Azedi, Elham Hoveizi, Jafar Ai and Mohammad Taghi Joghataei, date: 01/20/2018, view: 607, Q&A: 0
    [Abstract] Human endometrial stem cell/stromal cells (hEnSCs) are isolated from endometrium or menstrual blood and are recognized as a valuable cell type in tissue engineering and cell therapy. Furthermore, hEnSCs, which have CD90 (a mesenchymal marker), CD105 (endoglin), CD44, CD146 (endometrial stem cell markers) and lack CD31 (Endothelial marker), CD34 ...
    FACS-based Isolation of Neural and Glioma Stem Cell Populations from Fresh Human Tissues Utilizing EGF Ligand
    Authors:  Jessica Tome-Garcia, Fiona Doetsch and Nadejda M. Tsankova, date: 12/20/2017, view: 1341, Q&A: 0
    [Abstract] Direct isolation of human neural and glioma stem cells from fresh tissues permits their biological study without prior culture and may capture novel aspects of their molecular phenotype in their native state. Recently, we demonstrated the ability to prospectively isolate stem cell populations from fresh human germinal matrix and glioblastoma ...
    Isolation of Primary Human Skeletal Muscle Cells
    Authors:  Janelle M. Spinazzola and Emanuela Gussoni, date: 11/05/2017, view: 1154, 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 and Purification of Schwann Cells from Spinal Nerves of Neonatal Rat
    Authors:  Jinkun Wen, Dandan Tan, Lixia Li and Jiasong Guo, date: 10/20/2017, view: 1760, Q&A: 0
    [Abstract] Primary cultured Schwann cells (SCs) are widely used in the investigation of the biology of SC and are important seed cells for neural tissue engineering. Here, we describe a novel protocol for harvesting primary cultured SCs from neonatal Sprague-Dawley (SD) rats. In the present protocol, dissociated SCs are isolated from the spinal nerves of ...
    Isolation and Separation of Epithelial CD34+ Cancer Stem Cells from Tgfbr2-deficient Squamous Cell Carcinoma
    Authors:  Heather A. McCauley and Géraldine Guasch, date: 09/05/2017, view: 1565, Q&A: 0
    [Abstract] Most epithelial tumors have been shown to contain cancer stem cells that are potentially the driving force in tumor progression and metastasis (Kreso and Dick, 2014; Nassar and Blanpain, 2016). To study these cells in depth, cell isolation strategies relying on cell surface markers or fluorescent reporters are essential, and the isolation ...