Intestinal Stem Cells: Methods and Protocols

✍ Scribed by Paloma Ordóñez-Morán

• Publisher: Springer US;Humana
• Year: 2020
• Tongue: English
• Leaves: 348
• Series: Methods in Molecular Biology 2171
• Edition: 1st ed.
• Category: Library

✦ Synopsis

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.

✦ Table of Contents

Front Matter ....Pages i-xv
Front Matter ....Pages 1-1
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
Immune-Mediated Specific Depletion of Intestinal Stem Cells (Stephen E. Sherman, Judith Agudo)....Pages 25-39
Analysis of Aged Dysfunctional Intestinal Stem Cells (Kodandaramireddy Nalapareddy, Hartmut Geiger)....Pages 41-52
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
Visualization of Stem Cell Niche by Fluorescence Lifetime Imaging Microscopy (Irina A. Okkelman, Jens Puschhof, Dmitri B. Papkovsky, Ruslan I. Dmitriev)....Pages 65-97
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
Autophagy Detection in Intestinal Stem Cells (Jumpei Asano, Taku Sato, Toshiaki Ohteki)....Pages 115-125
Front Matter ....Pages 127-127
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
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
Front Matter ....Pages 169-169
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
Primary Intestinal Epithelial Organoid Culture (Tomohiro Mizutani, Hans Clevers)....Pages 185-200
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
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
Direct Lineage Reprogramming of Mouse Fibroblasts to Acquire the Identity of Fetal Intestine-Derived Progenitor Cells (Shizuka Miura, Atsushi Suzuki)....Pages 231-236
Single-Molecule RNA FISH in Whole-Mount Organoids (Costanza Borrelli, Andreas E. Moor)....Pages 237-247
Specific Gene Expression in Lgr5+ Stem Cells by Using Cre-Lox Recombination (Pierre Dessen, Joerg Huelsken, Paloma Ordóñez-Morán)....Pages 249-255
Generating and Utilizing Murine Cas9-Expressing Intestinal Organoids for Large-Scale Knockout Genetic Screening (Hossein Kashfi, Nicholas Jinks, Abdolrahman S. Nateri)....Pages 257-269
Front Matter ....Pages 271-271
Mouse Model for Sporadic Mutation of Target Alleles to Understand Tumor Initiation and Progression and Stem Cell Dynamics (Theresa N. Nguyen, Elise C. Manalo, Taryn E. Kawashima, Jared M. Fischer)....Pages 273-284
Hemagglutinating Virus of Japan Envelope (HVJ-E)-Guided Gene Transfer to the Intestinal Epithelium (Masamichi Imajo)....Pages 285-291
An Intrasplenic Injection Model for the Study of Cancer Stem Cell Seeding Capacity (Caroline Dafflon, Albert Santamaría-Martínez, Paloma Ordóñez-Morán)....Pages 293-302
Organoid Derivation and Orthotopic Xenotransplantation for Studying Human Intestinal Stem Cell Dynamics (Shinya Sugimoto, Masayuki Fujii, Toshiro Sato)....Pages 303-320
Advanced Colorectal Cancer Orthotopic Patient-Derived Xenograft Models for Cancer and Stem Cell Research (Irene Chicote, Juan Antonio Cámara, Héctor G. Palmer)....Pages 321-329
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 et al.)....Pages 331-346
Back Matter ....Pages 347-350

✦ Subjects

Life Sciences; Stem Cells; Cancer Research; Anatomy; Cell Biology