Osteil Pierre (ed.) Epiblast Stem Cells: Methods and Protocols

Humana Press, 2022. — 301 p. — (Methods in Molecular Biology 2490). — ISBN 978-1-0716-2280-3.This book aims to deliver the most up-to-date protocols in using epiblast stem cells (EpiSC) to answer critical questions on mammalian development. Beginning with a section on the spectrum of mouse primed pluripotent stem cells, the volume continues with chapters on toolkits to characterize and study primed stem cells, as well as primed stem cells to engineer gastrulation models. Written for the highly successful Method in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Epiblast Stem Cells: Methods and Protocols serves as an ideal guide to overcome the reproducibility problem by gathering detailed state-of-the-art protocols to perform comprehensive analyses, enabling the reader to opt for the appropriate cell type to model the epiblast.The Spectrum of Mouse Primed Pluripotent Stem Cells
Establishment of Mouse Epiblast Stem Cells
L-Proline Supplementation Drives Self-Renewing Mouse Embryonic Stem Cells to a Partially Primed Pluripotent State: The Early Primitive Ectoderm-Like Cell
Generation of Epiblast-Like Cells
Toolkits to Characterize and Study Primed Stem Cells
Identification and Visualization of Protein Expression in Whole Mouse Embryos by Immunofluorescence
Small Interfering RNA (siRNA) Transfection in Epiblast Stem Cells
Measuring Endocytosis and Endosomal Uptake at Single Cell Resolution
3D Immunofluorescent Image Colocalization Quantification in Mouse Epiblast Stem Cells
Flow Cytometric Characterization of Pluripotent Cell Protein Markers in Naïve, Formative, and Primed Pluripotent Stem Cells
Exploring Chromatin Accessibility in Mouse Epiblast Stem Cells with ATAC-Seq
A Reproducible and Dynamic Workflow for Analysis and Annotation of scRNA-Seq Data
Complete Transcriptome Analysis by 5′-End Single-Cell RNA-Seq with Random Priming
Global Proteomic Profiling of Embryonic Stem Cells Using iTRAQ Isobaric Tags with LC-MS/MS Quantification
Comprehensive and Comparative Structural Glycome Analysis in Mouse Epiblast-like Cells
Primed Stem Cells to Engineer Gastrulation Model
Generation of Gastruloids from Epiblast-Like Cells
Definitive Endoderm from EpiSC Aggregates in Matrigel
In Vitro Differentiation of Murine Embryonic Stem Cells (ESCs) into Primordial Germ Cell-like Cells (PGCLCs)
Primed Stem Cells to Engineer Gastrulation Model
Differentiation of Human-Induced Pluripotent Stem Cells (hiPSCs) into Human Primordial Germ Cell-like Cells (hPGCLCs) In Vitro
Differentiation of EpiLCs on Micropatterned Substrates Generated by Micro-Contact Printing
Grafting of Epiblast Stem Cell into the Epiblast and Whole-Embryo Imaging to Unveil Lineage Competence
Modeling Epiblast Shape in Implanting Mammalian Embryos