Ordóñez-Morán Intestinal Stem Cells

Methods and Protocols
2. Auflage 2027
ISBN: 978-1-0716-5412-5
Verlag: Springer US
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, 312 Seiten

Reihe: Springer Protocols

This second volume provides new and up-to-date methods of the intestinal stem cell field. Chapters guide readers through techniques, single-cell transcriptional profiling method, the isolation of intestinal crypts to generate, establish 3D organoids, animal models of gastrointestinal cancer, and examples of the use of methods for studying intestinal tumor-initiating cells. Written for the highly successful 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,  aims to provide comprehensive and easy to follow protocols.

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Ordóñez-Morán, Paloma

Inhaltsverzeichnis

Intestinal stem cells: regulation, plasticity, and therapeutic Potential.- In silico single-cell framework for modelling intestinal stem and transit-amplifying progenitor cells dynamics.- Using in silico pseudobulk differential expression and trajectory analysis of single cell RNA sequencing to reveal stem cell state transitions in colonic inflammation.- Cross-species translation of drug-Induced toxicity in the small intestine using mathematical modelling.- A computational framework to resolve stem cell gene dynamics during intestinal inflammation.- Animal-free intestinal epithelial 2D “Organoid-Sheet” cultures using invasion.- Primary cultures of foetal epithelial progenitors.- An integrated workflow for culturing and AI-based profiling of mouse intestinal epithelium.- Updated protocols for 3D and 2D culture and histological analysis of intestinal organoids.- Rapid and efficient genetic manipulation of intestinal organoids under transient two-dimensional culture conditions.- Human small intestinal organoids differentiation protocols towards all mature epithelial lineages.- Mass spectrometry-based proteomics of intestinal organoids.- Organoid culture model for the study of adult slow-cycling putative intestinal stem cells.- Intestinal organoids culture in synthetic hydrogels.- Live Fluorescence Lifetime Imaging Microscopy (FLIM) of the epithelial polarity and cell-specific segmentation in the small intestinal organoids.- Whole-mount immunostaining of human intestinal organoids.- Monitoring calcium dynamics in pig intestinal organoids with FLIM and FRET.- Methods for processing stem cell-derived organoids for histological and immunofluorescence staining.- Stemness in colon cancer.- Deep insights without clearing: an optimized protocol for patient-derived whole-organoid immunofluorescence imaging.- Comprehensive isolation strategy for intestinal epithelial and fibroblast cells from both healthy and tumour human tissues.- A tissue explant model for colon adenocarcinoma.- A straightforward and reliable mouse model of colorectal cancer by orthotopic transplantation.