Protocols in Current Issue
    Isolation and Culture of Murine Hepatic Stellate Cells
    Authors:  Rucha V. Modak and Dietmar M. Zaiss, date: 11/05/2019, view: 138, Q&A: 0
    [Abstract] Hepatic stellate cells (HSCs), alternatively known as liver pericytes, can differentiate into myofibroblasts and secrete extra-cellular matrix components, thereby promoting wound healing and fibrosis. Studying HSCs can provide insights into the pathological mechanisms governing these processes. HSC isolation methods typically comprise of enzymatic ...
    High Throughput Traction Force Microscopy for Multicellular Islands on Combinatorial Microarrays
    Authors:  Ian C. Berg and Gregory H. Underhill, date: 11/05/2019, view: 113, Q&A: 0
    [Abstract] The composition and mechanical properties of the cellular microenvironment along with the resulting distribution of cellular devolved forces can affect cellular function and behavior. Traction Force Microscopy (TFM) provides a method to measure the forces applied to a surface by adherent cells. Numerous TFM systems have been described in ...
    Lipid-exchange Rate Assay for Lipid Droplet Fusion in Live Cells
    Authors:  Jia Wang, Boon Tin Chua, Peng Li and Feng-Jung Chen, date: 07/20/2019, view: 751, Q&A: 0
    [Abstract] Lipid droplets (LDs) are central organelles in maintaining lipid homeostasis. Defective LD growth often results in the development of metabolic disorders. LD fusion and growth mediated by cell death–inducing DNA fragmentation factor alpha (DFFA)-like effector (CIDE) family proteins are crucial for various biological processes including unilocular ...
    Extraction and Analysis of Pan-metabolome Polar Metabolites by Ultra Performance Liquid Chromatography–Tandem Mass Spectrometry (UPLC-MS/MS)
    Authors:  Dania M. Malik, Seth Rhoades and Aalim Weljie, date: 02/05/2018, view: 3329, Q&A: 0
    [Abstract] Modern triple quadrupole mass spectrometers provide the ability to detect and quantify a large number of metabolites using tandem mass spectrometry (MS/MS). Liquid chromatography (LC) is advantageous, as it does not require derivatization procedures and a large diversity in physiochemical characteristics of analytes can be accommodated through a ...
    Generation of Chemically Induced Liver Progenitors (CLiPs) from Rat Adult Hepatocytes
    Authors:  Takeshi Katsuda, Kazunori Hosaka and Takahiro Ochiya, date: 01/20/2018, view: 4313, 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: 5356, 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: 7293, 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: 6712, 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. ...
    Determination of the H+-ATP Synthase and Hydrolytic Activities
    Authors:  Javier García-Bermúdez, Cristina Nuevo-Tapioles and José M. Cuezva, date: 08/20/2016, view: 5055, Q&A: 0
    [Abstract] The H+-ATP synthase of the inner mitochondrial membrane utilizes the proton gradient generated by the respiratory chain to synthesize ATP. Under depolarizing conditions, it can function in reverse by hydrolyzing ATP to generate a proton gradient. The protocols presented here allow the facile determination of both the synthetic and ...
    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: 6606, 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 ...

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