Epiblast stem cells : methods and protocols / edited by Pierre Osteil. -- New York, NY : Humana Press, 2022. – (58.17/M592/v.2490)

Contents

Preface
Contributors
PART I  THE SPECTRUM OF MOUSE PRIMED PLURIPOTENT STEM CELLS
1  Establishment of Mouse Epiblast Stem Cells
2  L-Proline Supplementation Drives Self-Renewing Mouse Embryonic Stem Cells to a Partially Primed Pluripotent State: The Early Primitive Ectoderm-Like Cell
3  Generation of Epiblast-Like Cells
PART II  TOOLKITS TO CHARACTERIZE AND STUDY PRIMED STEM CELLS
4  Identification and Visualization of Protein Expression in Whole Mouse Embryos by Immunofluorescence
5  Small Interfering RNA (siRNA) Transfection in Epiblast Stem Cells
6  Measuring Endocytosis and Endosomal Uptake at Single Cell Resolution
7  3D Immunofluorescent Image Colocalization Quantification in Mouse Epiblast Stem Cells
8  Flow Cytometric Characterization of Pluripotent Cell Protein Markers in Naive, Formative, and Primed Pluripotent Stem Cells
9  Exploring Chromatin Accessibility in Mouse Epiblast Stem Cells with ATAC-Seq
10  A Reproducible and Dynamic Workflow for Analysis and Annotation of scRNA-Seq Data
11  Complete Transcriptome Analysis by 5'-End Single-Cell RNA-Seq with Random Priming
12  Global Proteomic Profiling of Embryonic Stem Cells Using iTRAQ Isobaric Tags with LC-MS/MS Quantification
13  Comprehensive and Comparative Structural Glycome Analysis in Mouse Epiblast-like Cells
PART III  PRIMED STEM CELLS TO ENGINEER GASTRULATION MODEL
14  Generation of Gastruloids from Epiblast-Like Cells
15  Definitive Endoderm from EpiSC Aggregates in Matrigel
16  In Vitro Differentiation of Murine Embryonic Stem Cells (ESCs) into Primordial Germ Cell-like Cells (PGCLCs)
17  Differentiation of Human-Induced Pluripotent Stem Cells (hiPSCs) into Human Primordial Germ Cell-like Cells (hPGCLCs) In Vitro
18  Differentiation of EpiLCs on Micropatterned Substrates Generated by Micro-Contact Printing
19  Grafting of Epiblast Stem Cell into the Epiblast and Whole-Embryo Imaging to Unveil Lineage Competence
20  Modeling Epiblast Shape in Implanting Mammalian Embryos
Index