Embryonic Stem Cell Protocols

This extensive volume explores areas of intense activity related to the very early commitment of stem cells to particular lineages and the progression of differentiation to mature cell stages. Research on embryonic stem cells continues to move very quickly, thus the kinds of studies continue to expand and diversify, and methodologies are continuously being refined and improved, which this book reflects. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions on 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.

Comprehensive and fully updated, Embryonic Stem Cell Protocols, Third Edition serves as an ideal reference for researchers investigating this rich area of study.

Includes cutting-edge methods and protocols in embryonic stem cell research Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt

Maximizing Clonal Embryonic Stem Cell Derivation by ERK Pathway Inhibition.- A Simple and Efficient Method of Slow Freezing for Human Embryonic Stem Cells and Induced Pluripotent Stem Cells.- Resolving Heterogeneity: Fluorescence Activated Cell Sorting of Dynamic Cell Populations from Feeder-Free Mouse Embryonic Stem Cell Culture.- Acquiring Ground State Pluripotency: Switching Mouse Embryonic Stem Cells from Serum/LIF Medium to 2i/LIF Medium.- Generation of Embryonic Stem Cells in Rabbits.- Gene Transfer into Pluripotent Stem Cells Via Lentiviral Transduction.- Imaging Pluripotency: Time-Lapse Analysis of Mouse Embryonic Stem Cells.- Expanding the Utility of FUCCI Reporters Using FACS-Based 'Omics Analysis.- Selection of Antibodies Interfering with Cell-Surface Receptor Signalling Using Embryonic Stem Cell Differentiation.- Delivering Antisense Morpholino Oligonucleotides to Target Telomerase Splice Variants in Human Embryonic Stem Cells.- Analysis of mRNA Translation Rate in Mouse Embryonic Stem Cells.- Generation and Purification of Definitive Endoderm Cells Generated from Pluripotent Stem Cells.- Definitive Endoderm Differentiation of Human Embryonic Stem Cells Combined with Selective Elimination of Undifferentiated Cells by Methionine Deprivation.- Selective Differentiation into Hematopoietic and Cardiac Cells from Pluripotent Stem Cells Based on the Expression of Cell Surface Markers.- Methods for Derivation of Multipotent Neural Crest Cells Derived from Human Pluripotent Stem Cells.- From Naive to Primed Pluripotency: In Vitro Conversion of Mouse Embryonic Stem Cells in Epiblast Stem Cells.- In Vitro Differentiation of Embryonic Stem Cells into Hematopoietic Lineage: Towards Erythroid Progenitor's Production.- Dopaminergic Differentiation of Human Embryonic Stem Cells on PA6-Derived Adipocytes.- Intracellular Calcium Measurements for Functional Characterization ofNeuronal Phenotypes.- In Vitro Differentiation of Pluripotent Stem Cells into Functional Islets Under 2D and 3D Culture Conditions and In Vivo Preclinical Validation of 3D Islets.- Generation of Corneal Keratocytes from Human Embryonic Stem Cells.- Maintenance, Transgene Delivery, and Pluripotency Measurement of Mouse Embryonic Stem Cells.- Generation of a Knockout Mouse Embryonic Stem Cell Line Using a Paired CRISPR/Cas9 Genome Engineering Tool.- Derivation of Neural Precursor Cells from Human Embryonic Stem Cells for DNA Methylomic Analysis.- 26S and PA28-20S Proteasome Activity in Cytosolic Extracts from Embryonic Stem Cells.- A Concise Protocol for siRNA-Mediated Gene Suppression in Human Embryonic Stem Cells.- Applying Shear Stress to Pluripotent Stem Cells.- Generating Inner Ear Organoids from Mouse Embryonic Stem Cells.- Differentiation of Adipocytes in Monolayer from Mouse Embryonic Stem Cells.- Pancreatic Differentiation from Murine Embryonic Stem Cells.- Osteogenic Differentiation from Embryonic Stem Cells.- Limbal Stromal Tissue Specific Stem Cells and Their Differentiation Potential to Corneal Epithelial Cells.