Description
This volume presents 30 state-of-the-art protocols and reviews to set up and apply primary hepatocyte cultures for research and screening purposes. The first part of the book focuses on the use of these particular liver-based in vitro models to study the different aspects of the hepatocyte life cycle, including cell growth, differentiation and cell death. The second part of the book is targeted towards the demonstration of the applicability of primary hepatocyte cultures, or liver-based in vitro models derived thereof, for functionality and toxicity testing. 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 and readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Protocols In In-Vitro Hepatocyte Research is intended for basic and applied researchers in the area of pharmacology and toxicology, both in academic and industrial settings. Part I: In Vitro Models of the Hepatocyte Life Cycle1. Isolation And Culture Of Mouse Hepatocytes: Gender-Specific Gene Expression Responses To Chemical Treatments Lyndsey Houseman, Mina Edwards, Ian R. Phillips, and Elizabeth A. Shephard 2. Cryopreservation of Hepatocytes Nicola J. Hewitt and Albert P. Li 3. Culture Conditions Promoting Hepatocyte Proliferation And Cell Cycle Synchronization Anne Corlu and Pascal Loyer 4. Immortalized Human Hepatic Cell Lines For In Vitro Testing And Research Purposes Eva Ramboer, Tamara Vanhaecke, Vera Rogiers, and Mathieu Vinken 5. Culture and Functional Characterization Of Human Hepatoma Hepg2 Cells Mara Teresa Donato, Laia Tolosa, and Mara Jos Gmez-Lechn 6. Establishment And Characterization Of An In Vitro Model Of Fas-Mediated Hepatocyte Cell Death Mathieu Vinken, Michal Maes, Sara Crespo Yanguas, Joost Willebrords, Tamara Vanhaecke, and Vera Rogiers 7. Serum-Free Directed Differentiation Of Human Embryonic Stem Cells To Hepatocytes Kate Cameron, Baltasar Lucendo-Villarin, Dagmara Szkolnicka, and David C. Hay 8. Human Skin-Derived Precursor Cells: Isolation, Expansion And Hepatic Differentiation Joery De Kock, Robim M. Rodrigues, Karolien Buyl, Tamara Vanhaecke, Vera Rogiers 9. Generation Of Hepatocytes From Pluripotent Stem Cells For Drug Screening and Developmental Modeling Richard L. Gieseck III, Ludovic Vallier, and Nicholas R.F. Hannan 10. Differentiation-Promoting Medium Additives For Hepatocyte Cultivation and Cryopreservation Varvara Gouliarmou, Olavi Pelkonen, and Sandra Coecke 11. Co-Culture and Long-Term Maintenance Of Hepatocytes Merav Cohen, Gahl Levy, and Yaakov Nahmias 12. Primary Hepatocytes In Sandwich Culture Janneke Keemink, Marlies Oorts, and Pieter Annaert 13. Establishing Liver Bioreactors For In Vitro Research Sofia P. Rebelo, Rita Costa, Marcos F.Q. Sousa, Catarina Brito, and Paula M. Alves 14. Epigenetic Modifications As Anti-Dedifferentiation Strategy For Primary Hepatocytes In Culture Jennifer Bolleyn, Joanna Fraczek, Vera Rogiers, and Tamara Vanhaecke 15. Transfection Of Primary Hepatocytes With Liver-Enriched Transcription Factors Using Adenoviral Vectors Marta Benet, Ramiro Jover, and Roque Bort Part II: In Vitro Methods to Probe Hepatocyte Functionality and Toxicity 16. Transcriptomics Of Hepatocytes Treated With Toxicants For Investigating Molecular Mechanisms Underlying Hepatotoxicity Vaibhav Shinde, Regina Stber, Harshal Nemade, Isaia Sotiriadou, Jrgen Hescheler, JanHengstler, and Agapios Sachinidis 17. Global Microrna Analysis In Primary Hepatocyte Cultures Julian Krauskopf, Almudena Espn Prez, Jos C. Kleinjans, and Theo M. de Kok 18. Mass Spectrometry-Based Proteomics For Relative Protein Quantification And Biomarker Identification In Primary Human Hepatocytes Lisa Dietz and Albert Sickmann 19. Targeted Metabolomics For Homocysteine-Related Metabolites In Primary Hepatocytes Irena Selicharov and Marek Kornek 20. Measurement Of Cytochrome P450 Enzyme Induction And Inhibition In Human Hepatoma Cells Robim M. Rodrigues, Joery De Kock, Tatyana Doktorova, Vera Rogiers, and Tamara Vanhaecke 21. Analysis Of Sinusoidal Drug Uptake Transporter Activities In Primary Human Hepatocytes Marc Le Ve, Elodie Jouan, Claire Denizot, Yannick Parmentier, and Olivier Fardel 22. Measurement Of Albumin Secretion As Functionality Test In Primary Hepatocyte Cultures Karolien Buyl, Joery De Kock, Jennifer Bolleyn, Vera Rogiers, and Tamara Vanhaecke 23. Measurement Of Blood Coagulation Factor Synthesis In Cultures Of Human Hepatocytes Stefan Heinz and Joris Braspenning 24. Functionality Testing Of Primary Hepatocytes In Culture By Measuring Urea Synthesis Jennifer Bolleyn, Vera Rogiers, and Tamara Vanhaecke 25. Assay Of Bile Acid Conjugation And Excretion In Human Hepatocytes Helene Johansson and Ewa C.S. Ellis 26. General Cytotoxicity Assessment By Means Of The MTT Assay Laia Tolosa, Mara Teresa Donato, and Mara Jos Gmez-Lechn 27. Measurement Of Apoptotic And Necrotic Cell Death In Primary Hepatocyte Cultures Michal Maes, Tamara Vanhaecke, Bruno Cogliati, Sara Crespo Yanguas, Joost Willebrords, Vera Rogiers, and Mathieu Vinken 28. Critical Factors In The Assessment Of Cholestatic Liver Injury in vitro Benjamin L. Woolbright and Hartmut Jaeschke 29. In vitro Cell Culture Models Of Hepatic Steatosis Gahl Levy, Merav Cohen, and Yaakov Nahmias 30. Assessment Of Liver Fibrotic Insults in vitro Luis Perea, Mar Coll, and Pau Sancho-Bru
