Despite political and ethical controversies surrounding the study of human embryonic stem (hES) cells, new freedoms in regard to using them for research has allowed interest to remain high in understanding the regulatory mechanisms of stem cell self-renewal, their differentiation along various lineages, and their potential use in regenerative medicine. In Human Embryonic Stem Cell Protocols, Second Edition, internationally respected researchers expand upon the popular first edition and describe in detail their most useful techniques for the molecular and cellular manipulation of these intriguing cells. This diverse collection of readily reproducible methods has been optimized for the derivation, characterization, and differentiation of hES cells, with special attention given to regenerative medicine applications. As a volume of the Methods in Molecular Biology(TM) series, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Human Embryo Culture and Assessment for the Derivation of Embryonic Stem Cells (ESC).- Derivation of Human Embryonic Stem Cell Lines from Vitrified Human Embryos.- Growth of Human Embryonic Stem Cells Using Derivates of Human Fibroblasts.- In Vitro Neural Differentiation of Human Embryonic Stem Cells Using a Low-Density Mouse Embryonic Fibroblast Feeder Protocol.- Optimization of Physiological Xenofree Molecularly Defined Media and Matrices to Maintain Human Embryonic Stem Cell Pluripotency.- Serum-Free and Feeder-Free Culture Expansion of Human Embryonic Stem Cells.- Single Cell Enzymatic Dissociation of Human Embryonic Stem Cells: A Straightforward, Robust, and Standardized Culture Method.- Monitoring Stemness in Long-Term hESC Cultures by Real-Time PCR.- Culture and Preparation of Human Embryonic Stem Cells for Proteomics-Based Applications.- A Two- and Three-Dimensional Approach for Visualizing Human Embryonic Stem Cell Differentiation.- Immunoflourescence and mRNA Analysis of Human Embryonic Stem Cells (hESCs) Grown Under Feeder-Free Conditions.- Study of Gap Junctions in Human Embryonic Stem Cells.- hESC Engineering by Integrase-Mediated Chromosomal Targeting.- Human Embryonic Stem Cell Differentiation on Periodontal Ligament Fibroblasts.- Generation of Neural Crest Cells and Peripheral Sensory Neurons from Human Embryonic Stem Cells.- Embryonic Stem Cells as a Model for Studying Melanocyte Development.- In Vitro Derivation of Chondrogenic Cells from Human Embryonic Stem Cells.- Vascular Differentiation of Human Embryonic Stem Cells in Bioactive Hydrogel-Based Scaffolds.- Differentiation of Neural Precursors and Dopaminergic Neurons from Human Embryonic Stem Cells.- Transplantation of Human Embryonic Stem Cells and Derivatives to the Chick Embryo.- Genetic Manipulation of Human Embryonic Stem Cells.- Genetic Manipulation of Human Embryonic Stem Cells in Serum and Feeder-Free Media.- Human-Induced Pluripotent Stem Cells: Derivation, Propagation, and Freezing in Serum- and Feeder Layer-Free Culture Conditions.