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Time-lapse Whole-field Fluorescence Imaging of Microglia Processes Motility in Acute Mouse Hippocampal Slices and Analysis

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Induction and Analysis of Anti-CD40-induced Colitis in Mice

Authors: Barbara Joyce-Shaikh, Daniel J. Cua and David Bauché, date: 02/05/2019, view: 1039, Q&A: 0

[Abstract] Colon inflammation or colitis affects more than 1 million people worldwide. Several pre-clinical models, including chemical-induced (i.e., DSS, TNBS) or pathogen-induced (i.e., Citrobacter rodentium) have been used to study mechanisms involved in the development and regulation of colitis. Anti-CD40 induced colitis model ...

Collagenase-based Single Cell Isolation of Primary Murine Brain Endothelial Cells Using Flow Cytometry

Authors: Cathrin J. Czupalla, Hanadie Yousef, Tony Wyss-Coray and Eugene C. Butcher, date: 11/20/2018, view: 2023, Q&A: 0

[Abstract] The brain endothelium is a highly specialized vascular structure that maintains the activity and integrity of the central nervous system (CNS). Previous studies have reported that the integrity of the brain endothelium is compromised in a plethora of neuropathologies. Therefore, it is of particular interest to establish a method that enables ...

Papain-based Single Cell Isolation of Primary Murine Brain Endothelial Cells Using Flow Cytometry

Authors: Hanadie Yousef, Cathrin J Czupalla, Davis Lee, Eugene C Butcher and Tony Wyss-Coray, date: 11/20/2018, view: 1723, Q&A: 0

[Abstract] Brain endothelial cells (BECs) form the integral component of the blood-brain barrier (BBB) which separates the systemic milieu from the brain parenchyma and protects the brain from pathogens and circulating factors. In order to study BEC biology, it was of particular interest to establish a method that enables researchers to investigate and ...

HIV_GKO: A Tool to Assess HIV-1 Latency Reversal Agents in Human Primary CD4⁺ T Cells

Authors: Emilie Battivelli and Eric Verdin, date: 10/20/2018, view: 1254, Q&A: 0

[Abstract] While able to suppress HIV replication in HIV infected individuals, combination antiretroviral therapy (ART) fails to eliminate viral latent reservoir, which consists in integrated transcriptional silenced HIV provirus. So far, identification of latently-infected cells has relied on activating cells to induce expression of HIV proteins which can ...

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: 3099, 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: 4135, 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: 3338, 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 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: 3634, 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 ...

Isolation and Analysis of Stromal Cell Populations from Mouse Lymph Nodes

Authors: Yannick O. Alexandre and Scott N Mueller, date: 08/20/2017, view: 4324, Q&A: 0

[Abstract] Our protocol describes a simple procedure for isolating stromal cells from lymph nodes (LN). LN are disrupted then enzymatically digested with collagenase and dispase to produce a single cell suspension that can be stained with fluorescently labelled antibodies and analysed by flow cytometry. This protocol will enable identification of ...

Efficient Generation of Multi-gene Knockout Cell Lines and Patient-derived Xenografts Using Multi-colored Lenti-CRISPR-Cas9

Authors: Lena Behrmann, Scott McComb, Júlia Aguadé-Gorgorió, Yun Huang, Mario Hermann, Pawel Pelczar, Adriano Aguzzi, Jean-Pierre Bourquin and Beat C. Bornhauser, date: 04/05/2017, view: 9713, Q&A: 0

[Abstract] CRISPR-Cas9 based knockout strategies are increasingly used to analyze gene function. However, redundancies and overlapping functions in biological signaling pathways can call for generating multi-gene knockout cells, which remains a relatively laborious process. Here we detail the application of multi-color LentiCRISPR vectors to simultaneously ...

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