Main 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.

Contents:

Part I: In Vitro Models of the Hepatocyte Life Cycle1. Isolation And Culture Of Mouse Hepatocytes: Gender-Specific Gene Expression Responses To Chemical TreatmentsLyndsey 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

Maria Teresa Donato, Laia Tolosa, and Maria Jose Gomez-Lechon

6. Establishment And Characterization Of An In Vitro Model Of Fas-Mediated Hepatocyte Cell Death

Mathieu Vinken, Michael 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 Stoeber, Harshal Nemade, Isaia Sotiriadou, Jurgen Hescheler, JanHengstler, and Agapios Sachinidis

17. Global Microrna Analysis In Primary Hepatocyte Cultures

Julian Krauskopf, Almudena Espin Perez, 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 Selicharova and Marek Korinek

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 Vee, 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, Maria Teresa Donato, and Maria Jose Gomez-Lechon

27. Measurement Of Apoptotic And Necrotic Cell Death In Primary Hepatocyte Cultures

Michael 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