Human Embryonic Stem Cell Protocols

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Springer Science & Business Media, 2006 - Science - 385 pages
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A comprehensive collection of diverse techniques for the molecular and cellular manipulation of human embryonic stem (hES) cells. These readily reproducible methods have been optimized for the derivation, characterization, and differentiation of hES cells, with special attention given to regenerative medicine applications. A companion CD provides color versions of all illustrations in the book. The protocols follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
 

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Contents

Blastocyst Culture for Deriving Human Embryonic Stem Cells
13
Propagation of Human Embryonic Stem Cells
23
Derivation and Maintenance of Human Embryonic Stem Cells
43
Isolation and Characterization of Human Embryonic Stem Cells
55
Routine Culture and Differentiation of Human
77
Culture of Human Embryonic Stem Cells on Human
91
Maintenance of Human Embryonic Stem Cells in Animal Serum
105
Manipulation of SelfRenewal in Human Embryonic Stem Cells
115
Derivation and Characterization of Hematopoietic Cells
179
Retroviral Transduction of Hematopoietic Progenitors
201
Genetic Manipulation of Human Embryonic Stem Cells
221
Designing Testing and Validating a Microarray
241
Microarray Approach to Identify the Signaling Network
267
Massively Parallel Signature Sequencing
285
Confrontation Cultures of Embryonic Stem Cells
313
A Novel Experimental Platform for Investigating Cancer
329

Derivation of Human Feeders for Prolonged Support
129
Transplantation of Human Embryonic Stem Cells
137
Derivation and Characterization of Neuronal Precursors
153
In Vitro Differentiation of Neural Precursors From Human
169
inHumanEmbryonicStemCells
347
Index
375
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