Intestinal Stem Cells

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.

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

Inhalt
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.<p