Protocols in Current Issue
    Isolation and Quantification of Mouse γδT-cells in vitro and in vivo
    Authors:  Isha Rana, Krithika Badarinath, Ravindra K. Zirmire and Colin Jamora, date: 09/05/2021, view: 1864, Q&A: 0

    The skin plays an important role in protecting the body from pathogens and chemicals in the external environment. Upon injury, a healing program is rapidly initiated and involves extensive intercellular communication to restore tissue homeostasis. The deregulation of this crosstalk can lead to abnormal healing processes and is the foundation of

    Isolation and Stimulation of Peritoneal Macrophages with Apoptotic Jurkat Cells to Produce IL-10
    Authors:  Mei Song and Xiaojing Ma, date: 12/20/2019, view: 4125, Q&A: 0
    [Abstract] Clearance of apoptotic cells by macrophages is critical to ensuring cellular homeostasis and suppression of autoimmunity. Macrophage recognition of apoptotic cells triggers an anti-inflammatory response, which is mediated by the release of IL-10, TGF-β etc. with concurrent inhibition of pro-inflammatory cytokines (such as TNFα, IL-12, ...
    Isolation and Long-term Cultivation of Mouse Alveolar Macrophages
    [Abstract] Alveolar macrophages (AM) are tissue-resident macrophages that colonize the lung around birth and can self-maintain long-term in an adult organism without contribution of monocytes. AM are located in the pulmonary alveoli and can be harvested by washing the lungs using the method of bronchoalveolar lavage (BAL). Here, we compared different ...
    Isolation of Highly Pure Primary Mouse Alveolar Epithelial Type II Cells by Flow Cytometric Cell Sorting
    Authors:  Meenal Sinha and Clifford A. Lowell, date: 11/20/2016, view: 22812, Q&A: 1
    [Abstract] In this protocol, we describe the method for isolating highly pure primary alveolar epithelial type II (ATII) cells from lungs of naïve mice. The method combines negative selection for a variety of lineage markers along with positive selection for EpCAM, a pan-epithelial cell marker. This method yields 2-3 x 106 ATII cells per mouse ...
    Isolation of Intestinal Mesenchymal Cells from Adult Mice
    Authors:  Vasiliki Koliaraki and George Kollias, date: 09/20/2016, view: 20989, Q&A: 0
    [Abstract] During the last 20 years intestinal mesenchymal cells (IMCs) have emerged as an important cell type that plays a central role in intestinal development and homeostasis, by providing both structural support and growth regulatory elements. IMCs also actively participate in wound healing responses, thus regulating pathologic conditions such as tissue ...
    Mouse Embryonic Fibroblast Cell Culture and Stimulation
    Authors:  Lian-Qun Qiu, Wi S. Lai, Deborah J. Stumpo and Perry J. Blackshear, date: 07/05/2016, view: 18941, Q&A: 0
    [Abstract] Culture of mouse embryonic fibroblast (MEF) cells represents a powerful system to test gene function due to their easy accessibility, rapid growth rates, and the possibility of a large number of experiments. Fibroblasts are a group of heterogeneous resident cells of mesenchymal origin that have various locations, diverse appearances and ...
    Whole-mount Enteroid Proliferation Staining
    Authors:  Caitlyn W. Barrett, Sarah P. Short, Yash A. Choksi and Christopher Shawn Williams, date: 06/20/2016, view: 10323, Q&A: 0
    [Abstract] Small intestinal organoids, otherwise known as enteroids, have become an increasingly utilized model for intestinal biology in vitro as they recapitulate the various epithelial cells within the intestinal crypt (Mahe et al., 2013; Sato et al., 2009). Assessment of growth dynamics within these cultures is an important ...
    Isolation and Culture of the Islets of Langerhans from Mouse Pancreas
    Authors:  Kate L Graham, Stacey Fynch, Evan G Pappas, Christina Tan, Thomas WH Kay and Helen E Thomas, date: 06/20/2016, view: 22192, Q&A: 1
    [Abstract] The islets of Langerhans are clusters of endocrine cells located within the pancreas. Insulin-producing beta cells are the major cell type within islets, with glucagon-producing alpha cells and somatostatin-producing delta cells the other major cell types. The beta cells are the target of immune-mediated destruction in type 1 diabetes (Graham et ...
    In vitro Differentiation of Murine Innate Lymphoid Cells from Common Lymphoid Progenitor Cells
    Authors:  Corey Seehus and Jonathan Kaye, date: 03/20/2016, view: 8686, Q&A: 0
    [Abstract] Subtypes of innate lymphoid cells (ILC), defined based on their cytokine secretion profiles and transcription factor expression, are important for host protection from pathogens and maintaining tissue homeostasis. ILCs develop from common lymphoid progenitors (CLP) in the bone marrow. Using the methods described here, we have previously shown that ...
    Generation and Maturation of Human Monocyte-derived DCs
    Authors:  Michela Spadaro, Monica Montone and Federica Cavallo, date: 08/05/2014, view: 22354, Q&A: 0
    [Abstract] Dendritic cells (DC) are antigen-presenting cells, which play a critical role in the regulation of the adaptive immune response. They act as a bridge between the innate and the adaptive immune systems. An approach to study their function and potentiality is to generate DC-like cells by culturing CD14+ monocyte-enriched peripheral blood ...

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