Human Embryonic Stem Cells Handbook

Stem cell research has achieved much, yet still faces many challenges. In this comprehensive handbook, experts provide step-by-step, proven protocols allowing both novices and experienced researchers to make a comparative assessment-and avoid common pitfalls.

Considerable advances have taken place since the initial isolation and characterization of human embryonic stem (HES) cells; however, significant challenges remain before their potential for restoration and regeneration processes in patients can be realized. Understanding the diversity amongst HES cell lines and realizing the ability to isolate lines with robust differentiation potential remain difficult. In the Human Embryonic Stem Cells Handbook, experts in the field provide an assortment of protocols that have been used by various laboratories around the world so as to allow both novices and experienced investigators to compare and contrast different approaches to HES cell isolation and characterization with the hope that, from these protocols, researchers might standardize approaches for HES cell biology. Written in the Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.

Authoritative and accessible, Human Embryonic Stem Cells Handbook serves as a valuable reference for scientists pursuing this vital field and its enormous potential.

Covers isolation characterization of numerous human embryonic stem cells as well as essential protocols in HES characterization Features detailed, cutting-edge methods and protocols Includes expert tips and key implementation advice for successful implementation

Inhalt

Establishment of New Lines of Human Embryonic Stem Cells: Evolution of the Methodology.- Successful Scale-Up and Quality Assessments of Human Embryonic Stem Cells for Cell Therapy: Challenges and Overview.- Human Embryonic Stem Cells Derived in Xeno-Free Conditions.- Procedures for Derivation and Characterization of Human Embryonic Stem Cells from Odense, Denmark.- Principles for Derivation of Human Embryonic Stem Cells.- Derivation and Maintenance of Undifferentiated Human Embryonic Stem Cells.- Establishment of hESC Lines from the Inner Cell Mass of Blastocyst-Stage Embryos and Single Blastomeres of 4-Cell Stage Embryos.- Analysis of LINE-1 Expression in Human Pluripotent Cells.- Characterization and Gene Expression Profiling of Five Human Embryonic Stem Cell Lines Derived in Taiwan.- Derivation of Human Embryonic Stem Cell Lines from Poor Quality Embryos.- Derivation, Expansion, and Characterization of Human Embryonic Stem Cell Lines from Aneuploid Embryos.- Mutated Human Embryonic Stem Cells for the Study of Human Genetic Disorders.- Single-Cell Enzymatic Dissociation of hESC Lines OxF1--OxF4 and Culture in Feeder-Free Conditions.- Protocol for Expansion of Undifferentiated Human Embryonic and Pluripotent Stem Cells in Suspension.- Suspension Bioreactor Expansion of Undifferentiated Human Embryonic Stem Cells.- Derivation, Propagation, and Characterization of Neuroprogenitors from Pluripotent Stem Cells (hESCs and hiPSCs).- Comparison of Neural Differentiation Potential of Human Pluripotent Stem Cell Lines Using a Quantitative Neural Differentiation Protocol.- Array-Comparative Genomic Hybridization Characterization of Human Pluripotent Stem Cells.- Chromatin Immunoprecipitation-Based Analysis of Gene Regulatory Networks Operative in Human Embryonic Stem Cells.- Analysis of the Methylome of Human Embryonic Stem Cells Employing Methylated DNA Immunoprecipitation Coupled to Next Generation Sequencing (MeDIP-seq).- Stable Isotope Labelling with Amino Acids in Cell Culture for Human Embryonic Stem Cell Proteomic Analysis.