Mammalia

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    Protocols in Current Issue
    Simultaneous Fluorescent Recordings of Extracellular ATP and Intracellular Calcium in Mammalian Cells
    Authors:  Nicholas Mikolajewicz and Svetlana V. Komarova, date: 05/20/2019, view: 753, Q&A: 0
    [Abstract] Extracellular ATP is a potent signaling molecule that stimulates intracellular calcium responses through purinergic (P2) receptors in mammalian cells. While extracellular ATP and intracellular calcium can be measured separately, simultaneous monitoring can offer additional insights into P2 receptor physiology. This protocol takes advantage of the ...
    Cell Microencapsulation and Cryopreservation with Low Molecular Weight Hyaluronan and Dimethyl Sulfoxide
    Authors:  H. Gurruchaga, L. Saenz del Burgo, G. Orive, R. M. Hernandez, J. Ciriza and J. L. Pedraz, date: 02/20/2019, view: 875, Q&A: 0
    [Abstract] Cryopreservation is commonly used for the storage of cells, tissues, organs or 3D cell-based products using ultra-low temperatures, which involves the immersion in liquid nitrogen for their long-term preservation. The cryopreservation of several microencapsulated cells is usually performed by the slow freezing with the dimethyl sulfoxide (DMSO) as ...
    Calvarial Bone Implantation and in vivo Imaging of Tumor Cells in Mice
    Authors:  Kyoko Hashimoto, Shingo Sato, Hiroki Ochi, Shu Takeda and Mitsuru Futakuchi, date: 02/05/2019, view: 1046, Q&A: 0
    [Abstract] Bone is one of common metastasis sites for many types of cancer. In bone metastatic microenvironment, tumor-bone interactions play a significant role in the regulation of osteolytic or osteoblastic bone metastasis. In order to investigate the direct interaction between tumor cells and bone tissue, it is essential to generate appropriate animal ...
    Cartilage Induction from Mouse Mesenchymal Stem Cells in High-density Micromass Culture
    [Abstract] Mesenchymal stem cells have the ability to differentiate into multiple lineages, including adipocytes, osteoblasts and chondrocytes. Mesenchymal stem cells can be induced to differentiate into chondrocytes in extracellular matrices, such as alginate or collagen gel. Mesenchymal stem cells in a cell pellet or micromass culture can be also induced ...
    Method for Studying the Effect of Gene Silencing on Bacterial Infection-induced ERK1/2 Signaling in Bone-marrow Derived Macrophages
    Authors:  Gaurav Kumar, Subhash B. Arya and Amit Tuli, date: 12/20/2018, view: 1383, Q&A: 0
    [Abstract] Macrophages are highly phagocytic cells that utilize various pathogen recognition receptors (PRRs) to recognize pathogen-associated molecular patterns (PAMPs). These PAMPs can be present within the microbe, such as bacterial CpG DNA, and are recognized by Toll-like receptor 9 (TLR9), a PRR present on the endosomal membrane of macrophages. PAMPs ...
    Using Stable Isotopes in Bone Marrow Derived Macrophage to Analyze Metabolism
    [Abstract] Using gas chromatography mass spectrometry (GC-MS) to analyze the citric acid cycle (CAC) and related intermediates (such as glutamate, glutamine, GABA, and aspartate) is an analytical approach to identify unexpected correlations between apparently related and unrelated pathways of energy metabolism. Intermediates can be as expressed as their ...
    In vitro Osteoclastogenesis Assays Using Primary Mouse Bone Marrow Cells
    Authors:  Xueqian Zhuang and Guohong Hu, date: 06/05/2018, view: 2770, Q&A: 1
    [Abstract] Osteoclasts are a group of bone-absorbing cells to degenerate bone matrix and play pivotal roles in bone growth and homeostasis. The unbalanced induction of osteoclast differentiation (osteoclastogenesis) in pathological conditions, such as osteoporosis, arthritis and skeleton metastasis of cancer, causes great pain, bone fracture, hypercalcemia ...
    Osteoblast Sorting and Intracellular Staining of CXCL12
    Authors:  Weihuan Wang, Gurnoor Majhail, Cui Lui and Lan Zhou, date: 05/20/2018, view: 2380, Q&A: 0
    [Abstract] Osteoblasts are bone marrow endosteum-lining niche cells playing important roles in the regulation of hematopoietic stem cells by secreting factors and cell adhesion molecules. Characterization of primary osteoblasts has been achieved through culture of outgrowth of collagenase treated bone. Immunophenotyping and flow-based analysis of long bone ...
    Differentiation of Myeloid-derived Suppressor Cells from Murine Bone Marrow and Their Co-culture with Splenic Dendritic Cells
    Authors:  Giada Mondanelli and Claudia Volpi, date: 09/20/2017, view: 4934, Q&A: 0
    [Abstract] Myeloid-derived suppressor cells (MDSCs) possess the ability to suppress the immune response, and to amplify the regulatory properties of other immune cells, i.e., dendritic cells. Here we describe a protocol in which MDSCs were differentiated from murine bone marrow cells, and CD11c+ dendritic cells were purified from murine ...
    In vitro Detection of Neutrophil Traps and Post-attack Cell Wall Changes in Candida Hyphae
    Authors:  Alex Hopke and Robert T. Wheeler, date: 04/05/2017, view: 4407, Q&A: 0
    [Abstract] In this protocol we describe how to visualize neutrophil extracellular traps (NETs) and fungal cell wall changes in the context of the coculture of mouse neutrophils with fungal hyphae of Candida albicans. These protocols are easily adjusted to test a wide array of hypotheses related to the impact of immune cells on fungi and the cell ...



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