Controversy surrounds the study of human embryonic stem cells. In this book, 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.

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

Provides easily followed detailed protocols

Covers numerous essential methods for culturing and analyzing human embryonic stem cells

Brings together different approaches to maintaining and differentiating human embryonic stem cells

Includes supplementary material: sn.pub/extras

Inhalt

1. Human Embryo Culture and Assessment for the Derivation of Embryonic Stem Cells-ESC A. Henry Sathananthan and Tiki Osianlis 2. Derivation of Human Embryonic Stem Cell Lines from Vitrified Human Embryos Teija T. Peura, Julia Schaft, and Tomas Stojanov 3. Growth of Human Embryonic Stem Cells Using Derivates of Human Fibroblasts Carmen Escobedo-Lucea and Miodrag Stojkovic 4. In Vitro Neural Differentiation of Human Embryonic Stem Cells Using a Low-Density Mouse Embryonic Fibroblast Feeder Protocol John 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. Sammak 5. Optimization of Physiological Xenofree Molecularly-Defined Media and Matrices to Maintain Human Embryonic Stem Cell Pluripotency Isabelle Peiffer, Romain Barbet, Antoinette Hatzfeld, Ma-Lin Li, and Jacques A. Hatzfeld 6. Serum-Free and Feeder-Free Culture Expansion of Human Embryonic Stem Cells Katherine E. Wagner and Mohan C. Vemuri 7. Single Cell Enzymatic Dissociation of Human Embryonic Stem Cells: A Straightforward, Robust, and Standardized Culture Method Catharina Ellerström, Johan Hyllner, and Raimund Strehl 8. Monitoring Stemness in Long-Term hESC Cultures by Real-Time PCR Amparo Galán and Carlos Simón 9. Culture and Preparation of Human Embryonic Stem Cells for Proteomics-Based Applications Charles C. King 10. A Two- and Three-Dimensional Approach for Visualizing Human Embryonic Stem Cell Differentiation Christian B. Brøchner, Peter S. Vestentoft, Niels Lynnerup, Claus Yding Andersen, and Kjeld Møllgård 11. Immunoflourescence and mRNA Analysis of Human Embryonic Stem Cells (hESCs) Grown Under Feeder-Free Conditions Aashir Awan, Roberto S. Oliveri, Pernille L. Jensen, Søren T. Christensen,and Claus Yding Andersen 12. Study of Gap Junctions in Human Embryonic Stem Cells Raymond C.B. Wong and Alice Pébay 13. hESC Engineering by Integrase-Mediated Chromosomal Targeting Ying Liu, Uma Lakshmipathy, Ali Ozgenc, Bhaskar Thyagarajan, Pauline Lieu, Andrew Fontes, Haipeng Xue, Kelly Sheyhing, Chad MacArthur, and Jonathan D. Chesnut 14. Human Embryonic Stem Cell Differentiation on Periodontal Ligament Fibroblasts Y. Murat Elçin, Bülend Inanç, and A. Eser Elçin 15. Generation of Neural Crest Cells and Peripheral Sensory Neurons from Human Embryonic Stem Cells Ronald S. Goldstein, Oz Pomp, Irina Brokhman, and Lina Ziegler 16. Embryonic Stem Cells as a Model for Studying Melanocyte Development Susan E. Zabierowski and Meenhard Herlyn 17. In Vitro Derivation of Chondrogenic Cells from Human Embryonic Stem Cells Wei Seong Toh, Eng Hin Lee, Mark Richards, and Tong Cao 18. Vascular Differentiation of Human Embryonic Stem Cells in Bioactive Hydrogel-Based Scaffolds Sharon Gerecht, Lino S. Ferreira, and Robert Langer 19. Differentiation of Neural Precursors and Dopaminergic Neurons from Human Embryonic Stem Cells Xiao-Qing Zhang and Su-Chun Zhang 20. Transplantation of Human Embryonic Stem Cells and Derivatives to the Chick Embryo Ronald S. Goldstein 21. Genetic Manipulation of Human Embryonic Stem Cells Silvina Epsztejn-Litman and Rachel Eiges 22. Genetic Manipulation of Human Embryonic Stem Cells in Serum and Feeder Free Media Stefan R. Braam, Chris Denning, and Christine L. Mummery 23. Human Induced Pluripotent Stem Cells: Derivation, Propagation, and Freezing in Serum- and Feeder Layer-Free Culture Conditions Hossein Baharvand, Mehdi Totonchi, Adeleh Taei, Ali Seifinejad, Nasser Aghdami,