Foreword ....................................................... ix
Preface ........................................................ xi
Acknowledgments .............................................. xiii
Editors and Contributors ....................................... xv
SECTION I INTRODUCTION
1 Introduction to Pluripotent Stem Cells: Biology and
Applications ................................................. 3
Maria Borowski and Gary S. Stein
2 Researching and Obtaining Established Stem Cell Lines ........ 9
Mai X. Luong, Kelly P. Smith, and Gary S. Stein
SECTION II LABORATORY GUIDE FOR HUMAN STEM CELL CULTURE:
PLURIPOTENT STEM CELL CULTURE
3 Basics of Cell Culture ...................................... 19
Alicia Allaire, Mai X. Luong, and Kelly P. Smith
4 The Stem Cell Laboratory .................................... 33
Alicia Allaire, Mai X. Luong, and Kelly P. Smith
5 Reagent Preparation ......................................... 41
Alicia Allaire, Mai X. Luong, and Kelly P. Smith
6 Preparation of Mouse Embryonic Fibroblasts for Culture of
Human Embryonic Stem Cells .................................. 59
Meng-Jiao Shi, Maria Borowski, and Kimberly Stencel
7 Thawing and Seeding of Frozen Mouse Embryonic Fibroblasts ... 87
Meng-Jiao Shi, Maria Borowski, and Kimberly Stencel
8 Thawing, Seeding, and Changing the Medium of Human
Embryonic Stem Cells ........................................ 95
Meng-Jiao Shi, Kimberly Stencel, and Maria Borowski
9 Passaging of Human Embryonic Stem Cells on Inactivated
Mouse Embryonic Fibroblast Plates .......................... 105
Meng-Jiao Shi, Kimberly Stencel, and Maria Borowski
10 Harvesting Human Embryonic Stem Cells for
Cryopreservation ........................................... 119
Meng-Jiao Shi, Kimberly Stencel, and Maria Borowski
11 Human Embryonic Stem Cell Culture on BD Matrigel™ with
mTeSR®1 Medium ............................................. 129
Meng-Jiao Shi, Kimberly Stencel, and Maria Borowski
SECTION III LABORATORY GUIDE FOR HUMAN STEM CELL
CULTURE: CHARACTERIZATION OF PLURIPOTENT STEM CELLS
12 Defining Pluripotency ...................................... 141
Kelly P. Smith and Mai X. Luong
13 Characterization of Human Embryonic Stem Cells by
Immunofluorescence Microscopy .............................. 149
Shirwin M, Pockwinse and Prachi N. Ghule
14 Preparation of Human Embryonic Stem Cell Samples for Flow
Cytometry .................................................. 159
Meng-Jiao Shi, Kimberly Stencel, and Maria Borowski
15 Characterization of Human Embryonic Stem Cell Gene
Expression Using Reverse Transcription Polymerase Chain
Reaction ................................................... 167
Alicia Allaire, Kelly P. Smith, and Mai X. Luong
16 Determining Pluripotency of Human Embryonic Stem Cells:
Embryoid Body Formation .................................... 191
Shirwin M. Pockwinse and Prachi N. Ghule
17 Characterization of Human Embryonic Stem Cells by
Cytogenetics: Karyotyping and Fluorescence In Situ
Hybridization .............................................. 199
Shirwin M. Pockwinse, Prachi N. Ghule and Anne Higgins
18 High-Resolution Chromatin Immunoprecipitation Assay ........ 211
Beatriz Pérez-Cadahia, Bojan Drobic, and James R. Davie
19 Assaying Pluripotency Via Teratoma Formation ............... 217
M. William Lensch and Tan A. Ince
SECTION IV PERSPECTIVES IN HUMAN STEM CELL TECHNOLOGIES
20 Genomic Analysis of Pluripotent Stem Cells ................. 237
David Lapointe
21 Proteomic Analysis of Human Pluripotent Cells .............. 245
Andy T.Y. Lau, Yan-Ming Xu, and Jen-Fu Chiu
22 Bioinformatics Strategies for Understanding Gene
Expression in Human Pluripotent Cells ...................... 257
Gustavo Glusman, Bruz Marzolf, Kai Wang, Ji-Hoon Cho,
Burak Kutlu, and Qiang Tian
23 Epigenetic Analysis of Pluripotent Cells ................... 273
Mojgan Rastegar, Geneviève P. Delcuve, and James
R. Davie
24 Differentiation of Pluripotent Stem Cells: An Overview ..... 289
Jeremy Micah Crook
25 Cellular Reprogramming: Current Technology,
Perspectives, and Generation of Induced Pluripotent
Cells ...................................................... 297
Tanja Dominko
SECTION V APPLICATIONS OF HUMAN EMBRYONIC STEM CELLS
26 Human Pluripotent Cells: The Biology of Pluripotency ....... 313
Li-Fang Chu and Thomas P. Zwaka
27 Human Pluripotent Cells for Regenerative Medicine:
Potential Applications for Regenerative Medicine ........... 327
Christopher C. Ford and Darreil N. Kotton
28 Therapeutic Applications of Human Embryonic Stem Cells ..... 339
Shi-Jiang Lu, Irina Klimanskaya, Edmund Mickunas, and
Robert Lanza
29 Human Stem Cells for Drug Screening, Toxicity,
Specificity, and Off-Target Effects ........................ 353
Arnaud Lacoste and Mark Burcin
30 Interfacing Stem Cells with Gene Therapy ................... 367
Christian Mueller and Terence R. Flotte
31 Tissue Engineering for Stem Cell Mediated Regenerative
Medicine ................................................... 377
Janet Zoldan, Thomas P. Kraehenbuehl, Abigail K.R.
Lytton-Jean, Robert S. Langer, and Daniel G. Anderson
Glossary ...................................................... 401
Index ......................................................... 413
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