Intestinal Stem Cells : Methods and Protocols [electronic resource]

  • Ordóñez-Morán, Paloma. editor
  • Humana
  • 2020
Intestinal Stem Cells : Methods and Protocols [electronic resource]


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.


Identification and Isolation of Human LGR5+ Cells Using an Antibody-Based Strategy
Immune-Mediated Specific Depletion of Intestinal Stem Cells
Analysis of Aged Dysfunctional Intestinal Stem Cells
Strategies for Measuring Induction of Fatty Acid Oxidation in Intestinal Stem and Progenitor Cells
Visualization of Stem Cell Niche by the Fluorescence Lifetime Imaging Microscopy
Generation and Quantitative Imaging of Enteroid Monolayers
Autophagy Detection in Intestinal Stem Cells
Single-Cell Transcriptional Profiling of the Intestinal Epithelium
Single-Cell Studies of Intestinal Stem Cell Heterogeneity during Homeostasis and Regeneration
Large-Scale Production of Recombinant Noggin and R-Spondin1 Proteins Required for the Maintenance of Stem Cells in Intestinal Organoid Cultures
Primary Intestinal Epithelial Organoid Culture
In Vivo Human PSC-Derived Intestinal Organoids to Study Stem Cell Maintenance
Generation of Knock-Out Gene-Edited Human Intestinal Organoids
Direct Lineage Reprogramming of Mouse Fibroblasts to Acquire the Identity of Fetal Intestine-Derived Progenitor Cells
Single Molecule RNA FISH in Whole-Mount Organoids
Specific Gene Expression in Lgr5+ Stem Cells by Using Cre-Lox Recombination
Generating and Utilizing Murine Cas9-Expressing Intestinal Organoids for Large Scale Knockout Genetic Screening
Mouse Model for Sporadic Mutation of Target Alleles to Understand Tumor Initiation and Progression and Stem Cell Dynamics
Hemagglutinating Virus of Japan Envelop (HVJ-E)-Guided Gene Transfer to the Intestinal Epithelium
An Intrasplenic Injection Model for the Study of Cancer Stem Cells Seeding Capacity
Organoid Derivation and Orthotopic Xenotransplantation for Studying Human Intestinal Stem Cell Dynamics
Advanced Colorectal Cancer Orthotopic Patient-Derived Xenograft Models for Cancer and Stem Cell Research
Modeling Colorectal Cancer Progression through Orthotopic Implantation of Organoids