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This volume details experimental protocols to study structure and function of hepatocytes in vivo and in vitro, and for characterizing heterogeneity and plasticity of hepatocytes in healthy and in injured liver. Chapters guide readers through separating hepatocyte subfractions, characterizing the heterogeneity, novel culture systems, hepatocyte-like cells from human pluripotent stem cells, analyzing drug metabolism, monitoring protein sorting, manipulating gene expression, and methods for expanding primary hepatoblastoma and identifying hepatocellular carcinoma subtypes. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.
Authoritative and cutting-edge, Hepatocytes: Methods and Protocols aims to provide techniques to further understand molecular mechanisms involved in the regulating hepatocyte functions and underlying liver diseases.
Sommario
Long-Term Expansion of Primary Hepatocyte Organoids.- Generation of in vivo traceable hepatocyte-like cells from human iPSCs.- Generation of hepatocyte organoids from human iPS cells.- Induction of bile canaliculi forming hepatocytes from human induced pluripotent stem cells.- Human hepatocyte transduction with adeno-associated virus vector.- Hepatocyte-directed delivery of Lipid-encapsulated small interfering RNA.- Co-culture model for hepatitis B virus infection using iPSC-derived liver progenitor cells and liver sinusoidal endothelial cells.- In vitro assay system to detect drug-induced bile acid-dependent cytotoxicity using hepatocytes.- Evaluation of urea cycle activity by metabolic flux analysis using mass spectrometry.- Live Cell Imaging of Biosynthetic Protein Transport in Hepatocytes.- Collagen sandwich culture of primary hepatocytes for image-based investigations.-Assessment of Hepatocyte Ploidy by Flow Cytometry.- Isolation of small hepatocyte-like progenitor cells from retrorsine/partial hepatectomy rat livers by laser micro-dissection.- Application of EdU-based DNA synthesis assay to measure hepatocyte proliferation in situ during liver regeneration.- Quantification of clonal expansion of hepatocytes in normal and injured liver.- Evaluation of SOX9-positive hepatocytes in human liver specimens and mature mouse hepatocytes.- Identification of genes regulating hepatocyte injury by a genome-wide CRISPR-Cas9 Screen.- Identification of subtypes of HCC using bioinformatics and the hepatocyte differentiation model.- 3D culture of primary patient-derived hepatoblastoma tumoroids.- Preparation of functional heaptocytes.
