Main 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: Methods and 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:

Preface...
Table of Contents...
Contributing Authors...

1. Histopathological Diagnosis of Nonalcoholic Steatohepatitis (NASH)
Shah Giashuddin and Mouyed Alawad

2. Generation of a Diet-Induced Mouse Model of Nonalcoholic Fatty Liver Disease
Amon Asgharpour and Arun J. Sanyal

3. Hydrodynamic Injection for Developing NASH Model
Haichuan Wang and Xin Chen

4. Measurement of Hepatic Lipids
Sarah Forman, Patrick Tso, and Min Liu

5. Measurement of Fatty Acid Oxidation in Mammalian Cells
Wei Wang, Yibao Ma, Tianhai He, Erin Mooney, Chunqing Guo, Xiang-Yang Wang, and Xianjun Fang

6. Measurement of In Vivo VLDL and Chylomicron Secretion
Siddhartha S. Ghosh, Jing Wang, and Shobha Ghosh

7. Isolation and Culture of Mouse Hepatocytes and Kupffer Cells (KC)
Rachel Mendoza, Indranil Banerjee, Saranya Chidambaranathan Reghupaty, Rajesh Yetirajam, Debashri Manna, and Devanand Sarkar

8. Mouse Bone Marrow Cell Isolation and Macrophage Differentiation
Rachel Mendoza, Indranil Banerjee, Debashri Manna, Saranya Chidambaranathan Reghupaty, Rajesh Yetirajam, and Devanand Sarkar

9. Purification and Isolation of Hepatic Stellate Cells
Sonia Lele, Seung Duk Lee, Devanand Sarkar, and Marlon F. Levy

10. Isolation of the Stromal Vascular Fraction from Adipose Tissue and Subsequent Differentiation into White or Beige Adipocytes
Jared S. Farrar and Rebecca K. Martin

11. Culture of Mouse Liver Ductal Organoids
Junkai Yan, Yunling Tai, and Huiping Zhou

12. Development of a Scalable Three-Dimensional Culture of Human Induced Pluripotent Stem Cells-Derived Liver Organoids
Giuseppe Pettinato, Lev T. Perelman, and Robert A. Fisher

13. Chromatin Immunoprecipitation Assay in Primary Mouse Hepatocytes and Mouse Liver
Simiao Xu, Yangyang Liu, and Ji Miao

14. Analysis of Liver Responses to Non-Alcoholic Steatohepatitis by mRNA-Sequencing
Christopher D. Green, Mikhail G. Dozmorov and Sarah Spiegel

15. Single-Cell RNA-Sequencing in NASH
Jana Hundertmark, Hilmar Berger, and Frank Tacke

16. Computational Pipeline for Next-Generation Sequencing (NGS) Studies in Genetics of NASH
Adrian Salatino, Silvia Sookoian, and Carlos J. Pirola

17. Metabolic Phenotyping in Mice with NASH using Indirect Calorimetry
Bin Ni, Shanshan Chen, Jared S. Farrar, and Francesco S. Celi

18. Analysis of Insulin Resistance in Nonalcoholic Steatohepatitis
Hyunbae Kim, Deqiang Zhang, Zhenfeng Song, Xin Tong, and Kezhong Zhang

19. Assessment of Lipotoxic Endoplasmic Reticulum (ER) Stress in Nonalcoholic Steatohepatitis (NASH)
Gopanandan Parthasarathy and Harmeet Malhi

20. Ex Vivo Dual-Hit Method for Inflammasome Activation in Liver
Debajyoti Das, Moumita Adak, and Partha Chakrabarti

21. In Vivo Analysis of Necrosis and Ferroptosis in Nonalcoholic Steatohepatitis (NASH)
Shinya Tsurusaki, Kazuko Kanegae, and Minoru Tanaka

22. Analysis of the Sphingolipidome in NAFLD
David Montefusco, Johana Lambert, Andrea Anderson, Jeremy Allegood, and L. Ashley Cowart

23. Bile Acid Profiling in Mouse Biofluids and Tissues
Bo Kong, Daniel Rizzolo, Rulaiha Taylor, and Grace L. Guo

24. Preparation and Characterization of a Liver Targeted, Poly(amidoamine) Based, Gene Delivery System
Kareem Ebeid, Sean M. Geary, and Aliasger K. Salem

25. Extraction of Exosomes and Exosomal miRNA from Mesenchymal Stem Cells
Zhao Lin, Andrew Wong, and Shiekh Alam

26. Generation of Adenovirus for In Vitro and In Vivo Studies of Hepatocytes Yangyang Liu, Simiao Xu, Yun Liu, Yashaswini Kelagere Mayige Gowda, and Ji Miao

Subject Index List...