Includes cutting-edge methods and protocols
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
Includes cutting-edge methods and protocols
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
Includes supplementary material: sn.pub/extras
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
María Teresa Donato, Laia Tolosa, and María José Gómez-Lechón
6. Establishment And Characterization Of An In Vitro Model Of Fas-Mediated Hepatocyte Cell Death
Mathieu Vinken, Michaël 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 Tranillegalscription Factors Using Adenoviral Vectors
Marta Benet, Ramiro Jover, and Roque Bort
Part II: In Vitro Methods to Probe Hepatocyte Functionality and Toxicity
16. Tranillegalscriptomics Of Hepatocytes Treated With Toxicants For Investigating Molecular Mechanisms Underlying Hepatotoxicity
Vaibhav Shinde, Regina Stöber, Harshal Nemade, Isaia Sotiriadou, Jürgen Hescheler, JanHengstler, and Agapios Sachinidis
17. Global Microrna Analysis In Primary Hepatocyte Cultures
Julian Krauskopf, Almudena Espín Pérez, 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 Selicharová and Marek Korínek
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 Vée, 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, María Teresa Donato, and María José Gómez-Lechón
27. Measurement Of Apoptotic And Necrotic Cell Death In Primary Hepatocyte Cultures
Michaël 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
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.
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.
Isolation And Culture Of Mouse Hepatocytes: Gender-Specific Gene Expression Responses To Chemical Treatments.- Cryopreservation of Hepatocytes.- Culture Conditions Promoting Hepatocyte Proliferation And Cell Cycle Synchronization.- Immortalized Human Hepatic Cell Lines For In Vitro Testing And Research Purposes.- Culture and Functional Characterization Of Human Hepatoma Hepg2 Cells.- Establishment And Characterization Of An In Vitro Model Of Fas-Mediated Hepatocyte Cell Death.- Serum-Free Directed Differentiation Of Human Embryonic Stem Cells To Hepatocytes.- Human Skin-Derived Precursor Cells: Isolation, Expansion And Hepatic Differentiation.- Generation Of Hepatocytes From Pluripotent Stem Cells For Drug Screening and Developmental Modeling.- Differentiation-Promoting Medium Additives For Hepatocyte Cultivation and Cryopreservation.- Co-Culture and Long-Term Maintenance Of Hepatocytes.- Primary Hepatocytes In Sandwich Culture.- Establishing Liver Bioreactors For In Vitro Research.- Epigenetic Modifications As Anti-Dedifferentiation Strategy For Primary Hepatocytes In Culture.- Transfection Of Primary Hepatocytes With Liver-Enriched Tranillegalscription Factors Using Adenoviral Vectors.- Tranillegalscriptomics Of Hepatocytes Treated With Toxicants For Investigating Molecular Mechanisms Underlying Hepatotoxicity.- Global Microrna Analysis In Primary Hepatocyte Cultures.- Mass Spectrometry-Based Proteomics For Relative Protein Quantification And Biomarker Identification In Primary Human Hepatocytes.- Targeted Metabolomics For Homocysteine-Related Metabolites In Primary Hepatocytes.- Measurement Of Cytochrome P450 Enzyme Induction And Inhibition In Human Hepatoma Cells.- Analysis Of Sinusoidal Drug Uptake Transporter Activities In Primary Human Hepatocytes.- Measurement Of Albumin Secretion As Functionality Test In Primary Hepatocyte Cultures.- Measurement Of Blood Coagulation Factor Synthesis In Cultures Of Human Hepatocytes.- Functionality Testing Of Primary HepatocytesIn Culture By Measuring Urea Synthesis.- Assay Of Bile Acid Conjugation And Excretion In Human Hepatocytes.- General Cytotoxicity Assessment By Means Of The MTT Assay.- Measurement Of Apoptotic And Necrotic Cell Death In Primary Hepatocyte Cultures.- Critical Factors In The Assessment Of Cholestatic Liver Injury in vitro.- In vitro Cell Culture Models Of Hepatic Steatosis.- Assessment Of Liver Fibrotic Insults in vitro.
Inhaltsverzeichnis
Isolation And Culture Of Mouse Hepatocytes: Gender-Specific Gene Expression Responses To Chemical Treatments.- Cryopreservation of Hepatocytes.- Culture Conditions Promoting Hepatocyte Proliferation And Cell Cycle Synchronization.- Immortalized Human Hepatic Cell Lines For In Vitro Testing And Research Purposes.- Culture and Functional Characterization Of Human Hepatoma Hepg2 Cells.- Establishment And Characterization Of An In Vitro Model Of Fas-Mediated Hepatocyte Cell Death.- Serum-Free Directed Differentiation Of Human Embryonic Stem Cells To Hepatocytes.- Human Skin-Derived Precursor Cells: Isolation, Expansion And Hepatic Differentiation.- Generation Of Hepatocytes From Pluripotent Stem Cells For Drug Screening and Developmental Modeling.- Differentiation-Promoting Medium Additives For Hepatocyte Cultivation and Cryopreservation.- Co-Culture and Long-Term Maintenance Of Hepatocytes.- Primary Hepatocytes In Sandwich Culture.- Establishing Liver Bioreactors For In Vitro Research.- Epigenetic Modifications As Anti-Dedifferentiation Strategy For Primary Hepatocytes In Culture.- Transfection Of Primary Hepatocytes With Liver-Enriched Tranillegalscription Factors Using Adenoviral Vectors.- Tranillegalscriptomics Of Hepatocytes Treated With Toxicants For Investigating Molecular Mechanisms Underlying Hepatotoxicity.- Global Microrna Analysis In Primary Hepatocyte Cultures.- Mass Spectrometry-Based Proteomics For Relative Protein Quantification And Biomarker Identification In Primary Human Hepatocytes.- Targeted Metabolomics For Homocysteine-Related Metabolites In Primary Hepatocytes.- Measurement Of Cytochrome P450 Enzyme Induction And Inhibition In Human Hepatoma Cells.- Analysis Of Sinusoidal Drug Uptake Transporter Activities In Primary Human Hepatocytes.- Measurement Of Albumin Secretion As Functionality Test In Primary Hepatocyte Cultures.- Measurement Of Blood Coagulation Factor Synthesis In Cultures Of Human Hepatocytes.- Functionality Testing Of Primary HepatocytesIn Culture By Measuring Urea Synthesis.- Assay Of Bile Acid Conjugation And Excretion In Human Hepatocytes.- General Cytotoxicity Assessment By Means Of The MTT Assay.- Measurement Of Apoptotic And Necrotic Cell Death In Primary Hepatocyte Cultures.- Critical Factors In The Assessment Of Cholestatic Liver Injury in vitro.- In vitro Cell Culture Models Of Hepatic Steatosis.- Assessment Of Liver Fibrotic Insults in vitro.
Klappentext
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.nAuthoritative 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.
Includes cutting-edge methods and protocols
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
Includes supplementary material: sn.pub/extras
