Intestinal Stem Cells: Methods and Protocols

ISBN-13: 9781071607497 / Angielski / Miękka / 2021 / 350 str.

Paloma Ordóñez-Morán

Spis treści

Part I: Characterization, Imaging, and Functional Assays

 

1. Identification and Isolation of Human LGR5+ Cells Using an Antibody-Based Strategy

            Michael K. Dame, Sha Huang, Durga Attili, Jason R Spence, and Justin A. Colacino

 

2. Immune-Mediated Specific Depletion of Intestinal Stem Cells

            Stephen E. Sherman and Judith Agudo

 

3. Analysis of Aged Dysfunctional Intestinal Stem Cells

            Kodandaramireddy Nalapareddy and Hartmut Geiger

 

4. Strategies for Measuring Induction of Fatty Acid Oxidation in Intestinal Stem and Progenitor Cells

            Chia-Wei Cheng, Omer H. Yilmaz, and Maria M. Mihaylova

 

5. Visualization of Stem Cell Niche by the Fluorescence Lifetime Imaging Microscopy

            Irina A. Okkelman, Jens Puschhof, Dmitri B. Papkovsky, and Ruslan I. Dmitriev

 

6. Generation and Quantitative Imaging of Enteroid Monolayers

            Laura E. Sanman, Ina W. Chen, Jake M. Bieber, Curtis A. Thorne, Lani F. Wu, and Steven J. Altschuler

 

7. Autophagy Detection in Intestinal Stem Cells

            Jumpei Asano, Taku Sato, and Toshiaki Ohteki

 

Part II: Single-Cell Transcriptional Profiling of the Small Intestinal Epithelium

 

8. Single-Cell Transcriptional Profiling of the Intestinal Epithelium

            Claudia Capdevila, Ruben I. Calderon, Erin C. Bush, Kismet Sheldon-Collins, Peter A. Sims, and Kelley S. Yan

 

9. Single-Cell Studies of Intestinal Stem Cell Heterogeneity during Homeostasis and Regeneration

            Maxim Norkin, Claudia Capdevila, Ruben I. Calderon, Tianhong Su, Maria Trifas, Paloma Ordóñez-Morán, and Kelley S. Yan

 

Part III: Organoids and Applications

 

10. Large-Scale Production of Recombinant Noggin and R-Spondin1 Proteins Required for the Maintenance of Stem Cells in Intestinal Organoid Cultures

            David L. Hacker and Paloma Ordóñez-Morán

 

11. Primary Intestinal Epithelial Organoid Culture

            Tomohiro Mizutani and Hans Clevers

 

12. In Vivo Human PSC-Derived Intestinal Organoids to Study Stem Cell Maintenance

            Simon Vales, Holly M. Poling, Nambirajan Sundaram, Michael A. Helmrath, and Maxime M. Mahe

 

13. Generation of Knock-Out Gene-Edited Human Intestinal Organoids

            Chathruckan Rajendra, Tomas Wald, Kevin Barber, Jason R. Spence, Faranak Fattahi, and Ophir D. Klein

 

14. Direct Lineage Reprogramming of Mouse Fibroblasts to Acquire the Identity of Fetal Intestine-Derived Progenitor Cells

            Shizuka Miura and Atsushi Suzuki

 

15. Single Molecule RNA FISH in Whole-Mount Organoids

            Costanza Borrelli and Andreas E. Moor

 

16. Specific Gene Expression in Lgr5⁺ Stem Cells by Using Cre-Lox Recombination

            Pierre Dessen, Joerg Huelsken, and Paloma Ordóñez-Morán

 

17. Generating and Utilizing Murine Cas9-Expressing Intestinal Organoids for Large Scale Knockout Genetic Screening

            Hossein Kashfi, Nicholas Jinks, and Abdolrahman Shams Nateri

 

Part IV: In Vivo Models

 

18. Mouse Model for Sporadic Mutation of Target Alleles to Understand Tumor Initiation and Progression and Stem Cell Dynamics

            Theresa Nguyen, Elise Manalo, Taryn Kawashima, and Jared M. Fischer

 

19. Hemagglutinating Virus of Japan Envelop (HVJ-E)-Guided Gene Transfer to the Intestinal Epithelium

            Masamichi Imajo

 

20. An Intrasplenic Injection Model for the Study of Cancer Stem Cells Seeding Capacity

            Caroline Dafflon, Albert Santamaría-Martínez, and Paloma Ordóñez-Morán

 

21. Organoid Derivation and Orthotopic Xenotransplantation for Studying Human Intestinal Stem Cell Dynamics

            Shinya Sugimoto, Masayuki Fujii, and Toshiro Sato

 

22. Advanced Colorectal Cancer Orthotopic Patient-Derived Xenograft Models for Cancer and Stem Cell Research

            Irene Chicote, Juan Antonio Cámara, and Héctor G. Palmer

 

23. Modeling Colorectal Cancer Progression through Orthotopic Implantation of Organoids

            Felipe de Sousa e Melo, Jonathan M. Harnoss, Noelyn Kljavin, Ryan Scott, Catherine Sohn, Kevin G. Leong, and Frederic J. de Sauvage

Blurb

This detailed book encapsulates the most up-to-date methods of the intestinal stem cell field and provides guidance on a variety of techniques for studying intestinal stem cells properties. Beginning with a section on in vitro techniques to study different aspects of the intestinal stem cell functions by innovative imaging and functional assays, the volume continues with chapters detailing the single-cell transcriptional profiling method, the isolation of intestinal crypts to generate and establish 3D organoids, as well as different animal models of gastrointestinal cancer and examples of the use of in vivo methods for studying intestinal tumor-initiating cells or cancer stem cells. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and state-of-the-art, Intestinal Stem Cells: Methods and Protocols aims to provide comprehensive and easy to follow protocols designed to be helpful to both seasoned researchers and newcomers to this dynamic field.