Mammalia

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    Protocols in Current Issue
    Generation of Chemically Induced Liver Progenitors (CLiPs) from Rat Adult Hepatocytes
    Authors:  Takeshi Katsuda, Kazunori Hosaka and Takahiro Ochiya, date: 01/20/2018, view: 3448, 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 ...
    Endpoint or Kinetic Measurement of Hydrogen Sulfide Production Capacity in Tissue Extracts
    Authors:  Christopher Hine and James R. Mitchell, date: 07/05/2017, view: 4776, Q&A: 0
    [Abstract] Hydrogen sulfide (H2S) gas is produced in cells and tissues via various enzymatic processes. H2S is an important signaling molecule in numerous biological processes, and deficiencies in endogenous H2S production are linked to cardiovascular and other health complications. Quantitation of steady-state H2S ...
    Determination of the Glycolysis and Lipogenesis in Culture of Hepatocytes
    Authors:  Pierre-Damien Denechaud and Luis Fajas, date: 11/05/2016, view: 6636, Q&A: 0
    [Abstract] Metabolic flux analyses are needed to provide insights into metabolic regulation that occurs in cells. The current protocol describes fast and reproducible methods for determining glycolysis and de novo lipogenesis of hepatocytes. Primary culture of hepatocytes is an ‘in vitro’ model useful to study liver glucose and lipid ...
    Experimental Liver Fibrosis and Intrasplenic Transplantation of CD45+ Bone Marrow Cells
    Authors:  Prakash Baligar, Sebanta Pokhrel and Asok Mukhopadhyay, date: 10/20/2016, view: 5860, Q&A: 1
    [Abstract] Liver fibrosis results from the excessive collagen deposition (collagen scar) by activated hepatic stellate cells (HpSCs), leading to the inhibition of normal liver regeneration and function. Fibrogenesis is a complex mechanism involving both the synthesis and degradation of matrix proteins by different cell types, mainly macrophages in the liver. ...
    Mouse Liver Mitochondria Isolation, Size Fractionation, and Real-time MOMP Measurement
    Authors:  Thibaud T. Renault, Mark P.A Luna-Vargas and Jerry E. Chipuk, date: 08/05/2016, view: 6038, Q&A: 0
    [Abstract] The mitochondrial pathway of apoptosis involves a complex interplay between dozens of proteins and lipids, and is also dependent on the shape and size of mitochondria. The use of cellular models in past studies has not been ideal for investigating how the complex multi-factor interplay regulates the molecular mechanisms of mitochondrial outer ...
    Isolating Liver Mitochondria by Differential Centrifugation
    Authors:  Ignacio Amigo, Javier Traba and Carlos B. Rueda, date: 05/20/2016, view: 8360, Q&A: 0
    [Abstract] In addition to methods aimed at the study of mitochondrial function in-situ, a full understanding of mitochondrial function requires their purification from cells or tissues under specific physiological or pathological conditions. This protocol illustrates a sequential procedure to obtain functional mitochondria with high yield from mice liver ...
    Clonal Culture of Mouse Liver Progenitor Cells
    Author:  Naoki Tanimizu, date: 10/20/2015, view: 7003, Q&A: 0
    [Abstract] Liver stem/progenitor cells (LPCs) are defined as bipotential cells differentiating into both hepatocytes and cholangiocytes. For analyzing their differentiation potential, clonal culture has been used for LPCs isolated by a cell sorter. In addition, we can use the culture to assess functions of target genes on differentiation potential of LPCs. ...
    Murine Liver Myeloid Cell Isolation Protocol
    [Abstract] In homeostasis, the liver is critical for the metabolism of nutrients including sugars, lipids, proteins and iron, for the clearance of toxins, and to induce immune tolerance to gut-derived antigens. These functions predispose the liver to infection by blood-borne pathogens, and to a variety of diseases ranging from toxin and medication-induced ...
    Isolation of ILC2 from Mouse Liver
    Authors:  Tamar Mchedlidze and Stefan Wirtz, date: 07/20/2014, view: 7829, Q&A: 0
    [Abstract] Group 2 innate lymphoid cells (ILC2) are a recently characterized cell population which lacks specific antigen receptors and contributes to immune responses at mucosal surfaces of lung and gut. Recently, we demonstrated that ILC2 expand in the context of chronic liver diseases in mice and contribute to pathology in an IL-33 dependent manner. Here, ...
    Murine in vivo CD8+ T Cell Killing Assay
    Authors:  Myoungjoo V. Kim, Weiming Ouyang, Will Liao, Michael Q. Zhang and Ming O. Li, date: 07/05/2014, view: 11859, Q&A: 0
    [Abstract] Antigen-specific killing ability of effector CD8+ T cells is critical for protective immunity against infection. Here, we describe in vivo cytotoxic T cell assay to examine effector function of antigen-specific CD8+ T cells. Mice infected with Listeria monocytogenes (L. monocytogenes) expressing ...



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