Contents

Preface v

Contributors x\

Guide to the Companion CD xv

1 Human Embryonic Stem Cells:

Isolation, Maintenance, and Differentiation Kursad Turksen and Tammy-Claire Troy 1

2 Blastocyst Culture for Deriving Human Embryonic Stem Cells AriffBongso 13

3 Propagation of Human Embryonic Stem Cells on Human Feeder Cells Mark Richards and Ariff Bongso 23

4 Derivation and Maintenance of Human Embryonic Stem Cells

Michal Amit and Joseph Itskovitz-Eldor 43

5 Isolation and Characterization of Human Embryonic Stem Cells Maisam Mitalipova and Gail Palmarini 55

6 Routine Culture and Differentiation of Human

Embryonic Stem Cells Jim McWhir, Davina Wojtacha, and Alison Thomson 77

7 Culture of Human Embryonic Stem Cells on Human and Mouse Feeder Cells Gautam Dravid, Holly Hammond, and Linzhao Cheng 91

8 Maintenance of Human Embryonic Stem Cells in Animal Serum-

and Feeder Layer-Free Culture Conditions Michal Amit and Joseph Itskovitz-Eldor 105

9 Manipulation of Self-Renewal in Human Embryonic Stem Cells

Through a Novel Pharmacological GSK-3 Inhibitor Noboru Sato and Ali H. Brivanlou 115

10 Derivation of Human Feeders for Prolonged Support of Human Embryonic Stem Cells Chui-Yee Fong and Ariff Bongso 129

11 Transplantation of Human Embryonic Stem Cells to the Chick Embryo Ronald S. Goldstein 137

12 Derivation and Characterization of Neuronal Precursors and Dopaminergic Neurons From Human Embryonic Stem Cells In Vitro Melissa Carpenter, Mahendra S. Rao, William J. Freed, and Xianmin Zeng 153

13 In Vitro Differentiation of Neural Precursors From Human

Embryonic Stem Cells Xue-Jun Li and Su-Chun Zhang 169

14 Derivation and Characterization of Hematopoietic Cells

From Human Embryonic Stem Cells Lisheng Wang, Chantal Cerdan, Pablo Menendez, and Mickie Bhatia 179

15 Retroviral Transduction of Hematopoietic Progenitors

Derived From Human Embryonic Stem Cells Pablo Menendez, Lisheng Wang, Chantal Cerdan, and Mickie Bhatia 201

16 Genetic Manipulation of Human Embryonic Stem Cells by Transfection

Rachel Eiges 221

17 Designing, Testing, and Validating a Microarray for Stem Cell Characterization Yongquan Luo, Bhaskar Bhattacharya, Amy X. Yang,

Raj K. Puri, and Mahendra S. Rao 241

18 Microarray Approach to Identify the Signaling Network

Responsible for Self-Renewal of Human Embryonic Stem Cells Noboru Sato and Ali H. Brivanlou 267

19 Massively Parallel Signature Sequencing Daixing Zhou, Mahendra S. Rao, Roger Walker,

Irina Khrebtukova, Christian D. Haudenschild, Takumi Miura, Shannon Decola, Eric Vermaas, Keith Moon, and Thomas J. Vasicek 285

20 Confrontation Cultures of Embryonic Stem Cells

With Multicellular Tumor Spheroids to Study Tumor-Induced Angiogenesis Maria Wartenberg, Andreas Finkensieper, J├╝rgen Hescheler, and Heinrich Sauer 313

21 A Novel Experimental Platform for Investigating Cancer

Growth and Anti-Cancer Therapy in a Human Tissue Microenvironment Derived From Human Embryonic Stem Cells Maty Tzukerman and Karl L. Skorecki 329

22 The Analysis of Mitochondria and Mitochondrial DNA in Human Embryonic Stem Cells Justin C. St. John, Alexandra Amaral, Emma Bowles,

Joao Facucho Oliveira, Rhiannon Lloyd, Mariana Freitas, Heather L. Gray, Christopher S. Navara, Gisela Oliveira, Gerald P. Schatten, Emma Spikings, and Joao Ramalho-Santos 347

Index

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