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Intestinal Stem Cells

Methods and Protocols

  • Book
  • © 2020

Overview

Editors:
  1. Paloma Ordóñez-Morán

    1. Division of Cancer and Stem Cells, School of Medicine, Biodiscovery Institute, Centre for Cancer Sciences, University of Nottingham, Nottingham, UK

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    You can also search for this editor in PubMed   Google Scholar

  • Includes cutting-edge techniques
  • Provides step-by-step detail essential for reproducible results
  • Contains key implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 2171)

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Table of contents (23 protocols)

  1. Front Matter

    Pages i-xv
    Download chapter PDF

  2. Characterization, Imaging, and Functional Assays

    1. Front Matter

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

      • Michael K. Dame, Sha Huang, Durga Attili, Jason R. Spence, Justin A. Colacino
      Pages 3-23

    3. Immune-Mediated Specific Depletion of Intestinal Stem Cells

      • Stephen E. Sherman, Judith Agudo
      Pages 25-39

    4. Analysis of Aged Dysfunctional Intestinal Stem Cells

      • Kodandaramireddy Nalapareddy, Hartmut Geiger
      Pages 41-52

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

      • Chia-Wei Cheng, Omer H. Yilmaz, Maria M. Mihaylova
      Pages 53-64

    6. Visualization of Stem Cell Niche by Fluorescence Lifetime Imaging Microscopy

      • Irina A. Okkelman, Jens Puschhof, Dmitri B. Papkovsky, Ruslan I. Dmitriev
      Pages 65-97

    7. Generation and Quantitative Imaging of Enteroid Monolayers

      • Laura E. Sanman, Ina W. Chen, Jake M. Bieber, Curtis A. Thorne, Lani F. Wu, Steven J. Altschuler
      Pages 99-113

    8. Autophagy Detection in Intestinal Stem Cells

      • Jumpei Asano, Taku Sato, Toshiaki Ohteki
      Pages 115-125

  3. Single-Cell Transcriptional Profiling of the Intestinal Epithelium

    1. Front Matter

      Pages 127-127
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    2. Single-Cell Transcriptional Profiling of the Intestinal Epithelium

      • Claudia Capdevila, Ruben I. Calderon, Erin C. Bush, Kismet Sheldon-Collins, Peter A. Sims, Kelley S. Yan
      Pages 129-153

    3. 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 et al.
      Pages 155-167

  4. Organoids and Applications

    1. Front Matter

      Pages 169-169
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    2. Large-Scale Production of Recombinant Noggin and R-Spondin1 Proteins Required for the Maintenance of Stem Cells in Intestinal Organoid Cultures

      • David L. Hacker, Paloma Ordóñez-Morán
      Pages 171-184

    3. Primary Intestinal Epithelial Organoid Culture

      • Tomohiro Mizutani, Hans Clevers
      Pages 185-200

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

      • Simon Vales, Holly M. Poling, Nambirajan Sundaram, Michael A. Helmrath, Maxime M. Mahe
      Pages 201-214

    5. Generation of Knockout Gene-Edited Human Intestinal Organoids

      • Chathruckan Rajendra, Tomas Wald, Kevin Barber, Jason R. Spence, Faranak Fattahi, Ophir D. Klein
      Pages 215-230

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

      • Shizuka Miura, Atsushi Suzuki
      Pages 231-236

    7. Single-Molecule RNA FISH in Whole-Mount Organoids

      • Costanza Borrelli, Andreas E. Moor
      Pages 237-247

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

      • Pierre Dessen, Joerg Huelsken, Paloma Ordóñez-Morán
      Pages 249-255
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Keywords

About this book

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.

Editors and Affiliations

  • Division of Cancer and Stem Cells, School of Medicine, Biodiscovery Institute, Centre for Cancer Sciences, University of Nottingham, Nottingham, UK

    Paloma Ordóñez-Morán

Bibliographic Information

  • Book Title: Intestinal Stem Cells

  • Book Subtitle: Methods and Protocols

  • Editors: Paloma Ordóñez-Morán

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-0716-0747-3

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC, part of Springer Nature 2020

  • Hardcover ISBN: 978-1-0716-0746-6Published: 24 July 2020

  • Softcover ISBN: 978-1-0716-0749-7Published: 25 July 2021

  • eBook ISBN: 978-1-0716-0747-3Published: 23 July 2020

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XV, 350

  • Number of Illustrations: 10 b/w illustrations, 67 illustrations in colour

  • Topics: Stem Cells, Cancer Research, Anatomy, Cell Biology

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