Full Description
This volume contains a comprehensive collection of laboratory protocols used by researchers to analyze varied aspects of non-alcoholic steatohepatitis (NASH). The chapters in this book cover topics such as methods for histological diagnosis of NASH; the purpose of generating an in vivo NASH model; protocols for isolating hepatocytes and Kupffer cells, bone marrow derived macrophages, and adipocytes; techniques to develop human pluripotent stem cells-derived liver organoids; single-cell and RNA-sequencing; and a description of how to extract exosomes and exosomal miRNAs from mesenchymal stem cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Thorough and practical, Non-Alcoholic Steatohepatitis: Methodsand Protocols is a valuable resource for new and experiences investigators studying NASH, and serves as an essential reference on NASH for basic and clinical researchers and students.
Contents
Histopathological Diagnosis of Nonalcoholic Steatohepatitis (NASH).- Generation of a Diet-Induced Mouse Model of Nonalcoholic Fatty Liver Disease.- Hydrodynamic Injection for Developing NASH Model.- Measurement of Hepatic Lipids.- Measurement of Fatty Acid Oxidation in Mammalian Cells.- Measurement of In Vivo VLDL and Chylomicron Secretion.- Isolation and Culture of Mouse Hepatocytes and Kupffer Cells (KC).- Mouse Bone Marrow Cell Isolation and Macrophage Differentiation.- Purification and Isolation of Hepatic Stellate Cells.- Isolation of the Stromal Vascular Fraction from Adipose Tissue and Subsequent Differentiation into White or Beige Adipocytes.- Culture of Mouse Liver Ductal Organoids.- Development of a Scalable Three-Dimensional Culture of Human Induced Pluripotent Stem Cells-Derived Liver Organoids.- Chromatin Immunoprecipitation Assay in Primary Mouse Hepatocytes and Mouse Liver.- Analysis of Liver Responsesto Non-Alcoholic Steatohepatitis by mRNA-Sequencing.- Single-Cell RNA-Sequencing in NASH.- Computational Pipeline for Next-Generation Sequencing (NGS) Studies in Genetics of NASH.- Metabolic Phenotyping in Mice with NASH using Indirect Calorimetry.- Analysis of Insulin Resistance in Nonalcoholic Steatohepatitis.- Assessment of Lipotoxic Endoplasmic Reticulum (ER) Stress in Nonalcoholic Steatohepatitis (NASH).- Ex Vivo Dual-Hit Method for Inflammasome Activation in Liver.- In Vivo Analysis of Necrosis and Ferroptosis in Nonalcoholic Steatohepatitis (NASH).- Analysis of the Sphingolipidome in NAFLD.- Bile Acid Profiling in Mouse Biofluids and Tissues.- Preparation and Characterization of a Liver Targeted, Poly(amidoamine) Based, Gene Delivery System.- Extraction of Exosomes and Exosomal miRNA from Mesenchymal Stem Cells.- Generation of Adenovirus for In Vitro and In Vivo Studies of Hepatocytes.
