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¿ 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.Comprehensive and cutting-edge, Human Embryonic Stem Cell Protocols, Second Edition offers both novice and expert researchers powerful tools essential to understanding the maintenance and differentiation of human embryonic stem cells, as well as their applications in regenerative medicine today.

Comprises a comprehensive guide to isolate culture and differentiate human embryonic stem cells in vitro

1. Human Embryo Culture and Assessment for the Derivation of Embryonic Stem Cells-ESCA. Henry Sathananthan and Tiki Osianlis2. Derivation of Human Embryonic Stem Cell Lines from Vitrified Human EmbryosTeija T. Peura, Julia Schaft, and Tomas Stojanov3. Growth of Human Embryonic Stem Cells Using Derivates of Human FibroblastsCarmen Escobedo-Lucea and Miodrag Stojkovic4. In Vitro Neural Differentiation of Human Embryonic Stem Cells Using a Low-Density Mouse Embryonic Fibroblast Feeder ProtocolJohn A. Ozolek, Esther P. Jane, James E. Esplen, Patti Petrosko, Amy K. Wehn, Teresa M. Erb, Sara E. Mucko, Lyn C. Cote, and Paul J. Sammak5. Optimization of Physiological Xenofree Molecularly-Defined Media and Matrices to Maintain Human Embryonic Stem Cell PluripotencyIsabelle Peiffer, Romain Barbet, Antoinette Hatzfeld, Ma-Lin Li, and Jacques A. Hatzfeld6. Serum-Free and Feeder-Free Culture Expansion of Human Embryonic Stem CellsKatherine E. Wagner and Mohan C. Vemuri7. Single Cell Enzymatic Dissociation of Human Embryonic Stem Cells: A Straightforward, Robust, and Standardized Culture MethodCatharina Ellerström, Johan Hyllner, and Raimund Strehl8. Monitoring Stemness in Long-Term hESC Cultures by Real-Time PCRAmparo Galán and Carlos Simón9. Culture and Preparation of Human Embryonic Stem Cells for Proteomics-Based ApplicationsCharles C. King10. A Two- and Three-Dimensional Approach for Visualizing Human Embryonic Stem Cell DifferentiationChristian B. Brøchner, Peter S. Vestentoft, Niels Lynnerup, Claus Yding Andersen, and Kjeld Møllgård11. Immunoflourescence and mRNA Analysis of Human Embryonic Stem Cells (hESCs) Grown Under Feeder-Free ConditionsAashir Awan, Roberto S. Oliveri, Pernille L. Jensen, Søren T. Christensen,and Claus Yding Andersen12. Study of Gap Junctions in Human Embryonic Stem CellsRaymond C.B. Wong and Alice Pébay13. hESC Engineering by Integrase-Mediated Chromosomal TargetingYing Liu, Uma Lakshmipathy, Ali Ozgenc, Bhaskar Thyagarajan, Pauline Lieu, Andrew Fontes, Haipeng Xue, Kelly Sheyhing, Chad MacArthur, and Jonathan D. Chesnut14. Human Embryonic Stem Cell Differentiation on Periodontal Ligament FibroblastsY. Murat Elçin, Bülend Inanç, and A. Eser Elçin15. Generation of Neural Crest Cells and Peripheral Sensory Neurons from Human Embryonic Stem CellsRonald S. Goldstein, Oz Pomp, Irina Brokhman, and Lina Ziegler16. Embryonic Stem Cells as a Model for Studying Melanocyte DevelopmentSusan E. Zabierowski and Meenhard Herlyn17. In Vitro Derivation of Chondrogenic Cells from Human Embryonic Stem CellsWei Seong Toh, Eng Hin Lee, Mark Richards, and Tong Cao18. Vascular Differentiation of Human Embryonic Stem Cells in Bioactive Hydrogel-Based ScaffoldsSharon Gerecht, Lino S. Ferreira, and Robert Langer19. Differentiation of Neural Precursors and Dopaminergic Neurons from Human Embryonic Stem CellsXiao-Qing Zhang and Su-Chun Zhang20. Transplantation of Human Embryonic Stem Cells and Derivatives to the Chick EmbryoRonald S. Goldstein21. Genetic Manipulation of Human Embryonic Stem CellsSilvina Epsztejn-Litman and Rachel Eiges22. Genetic Manipulation of Human Embryonic Stem Cells in Serum and Feeder Free MediaStefan R. Braam, Chris Denning, and Christine L. Mummery23. Human Induced Pluripotent Stem Cells: Derivation, Propagation, and Freezing in Serum- and Feeder Layer-Free Culture ConditionsHossein Baharvand, Mehdi Totonchi, Adeleh Taei, Ali Seifinejad, Nasser Aghdami