Main description:

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.

Contents:

1. Long-Term Expansion of Primary Hepatocyte Organoids

Thomas A. Kluiver, Lianne J. Kraaier, and Weng Chuan Peng

2. Generation of in vivo traceable hepatocyte-like cells from human iPSCs

Candice Ashmore-Harris and Gilbert O. Fruhwirth

3. Generation of hepatocyte organoids from human iPS cells

Giuseppe Pettinato

4. Induction of bile canaliculi forming hepatocytes from human induced pluripotent stem cells

Lavinija Matakovic, Arend W. Overeem, Karin Klappe, and Sven C. D. van IJzendoorn

5. Human hepatocyte transduction with adeno-associated virus vector

Zhenwei Song, Wenwei Shao, Liujiang Song1, Xieolei Pei, and Chengwen Li

6. Hepatocyte-directed delivery of Lipid-encapsulated small interfering RNA

Laura Moran, Marius Maximilian Woitok, Matthias Bartneck, and Francisco Javier Cubero

7. Co-culture model for hepatitis B virus infection using iPSC-derived liver progenitor cells and liver sinusoidal endothelial cells

Misao Himeno, Shin-Wei Chen, and Taketomo Kido

8. In vitro assay system to detect drug-induced bile acid-dependent cytotoxicity using hepatocytes

Akinori Takemura, and Kosei Itoh

9. Evaluation of urea cycle activity by metabolic flux analysis using mass spectrometry

Yasuharu Ueno, Takuji Maeda, Satoshi Okamoto, and Hideki Taniguchi

10. Live Cell Imaging of Biosynthetic Protein Transport in Hepatocytes

Francisco Lazaro-Dieguez and Anne Musch

11. Collagen sandwich culture of primary hepatocytes for image-based investigations

Dong Fu

12. Assessment of Hepatocyte Ploidy by Flow Cytometry

Yinhua Jin, Teni Anbarchian, and Roel Nusse

13. Isolation of small hepatocyte-like progenitor cells from retrorsine/partial hepatectomy rat livers by laser micro-dissection

Norihisa Ichinohe, Naoki Tanimizu, and Toshihiro Mitaka

14. Application of EdU-based DNA synthesis assay to measure hepatocyte proliferation in situ during liver regeneration

Amit Ghosh, Awais Ullah Ihsan, Madhuparna Nandi, Maryse Cloutier, Md Gulam Musawwir Khan, Sheela Ramanathan and Subburaj Ilangumaran

15. Quantification of clonal expansion of hepatocytes in normal and injured liver

Feng Chen

16. Evaluation of SOX9-positive hepatocytes in human liver specimens and mature mouse hepatocytes

Yuji Suzuki, Tokio Sasaki, Keisuke Kakisaka, Hiroaki Abe, and Yasuhiro Takikawa

17. Identification of genes regulating hepatocyte injury by a genome-wide CRISPR-Cas9 Screen

Katherine Shortt and Daniel P. Heruth

18. Identification of subtypes of HCC using bioinformatics and the hepatocyte differentiation model

Wei Cheng, Yan Zhang, and Ming Liu

19. 3D culture of primary patient-derived hepatoblastoma tumoroids

Peng V. Wu and Roel Nusse

20. Preparation of functional heaptocytes

Ayumu Okumura and Naoki Tanimizu