Human Pluripotent Stem Cells: Methods and Protocols

Section I. Laboratory Essentials

1. The Stem Cell Laboratory: Design, Equipment, and Oversight

Robin L. Wesselschmidt and Philip H. Schwartz

2. Stem Cell Banks: Preserving Cell Lines, Maintaining Genetic Integrity, and Advancing Research

Lesley Young, Lyn Healy, and Glyn N. Stacey

Section II. Derivation

3. Embryonic Stem Cell Derivation from Human Embryos

Paul Lerou

4. Derivation of Human Parthenogenetic Stem Cell Lines

Nikolay Turovets, Andrey Semechkin, Leonid Kuzmichev, Jeffrey Janus, Larissa

Agapova, and Elena Revazova

5. Generation of Induced Pluripotent Stem Cell Lines from Human Fibroblasts via Retroviral Gene Transfer

Justine D. Miller and Thorsten M. Schlaeger

6. Derivation of Induced Pluripotent Stem Cells by Lentiviral Transduction

Hubert E. Nethercott, David J. Brick, and Philip H. Schwartz

7. Transgene-free Production of Pluripotent Stem Cells using PiggyBac Transposons

Knut Woltjen, Riikka Hämäläinen, Mark Kibschull, Maria Mileikovsky, and Andras Nagy

Section III. Growth, Maintenance and Expansion

8. Traditional Human Embryonic Stem Cell Culture

Philip H. Schwartz, David J. Brick, Hubert E. Nethercott, and Alexander E. Stover

9. Xeno-Free Culture of Human Pluripotent Stem Cells

Rosita Bergström, Susanne Ström, Frida Holm, Anis Feki, and Outi Hovatta

10. Adaptation of Human Pluripotent Stem Cells to Feeder-Free Conditions in Chemically-Defined Medium with Enzymatic Single-Cell Passaging

Alexander E. Stover and Philip H. Schwartz

11. GMP Scale-Up and Banking of Pluripotent Stem Cells for Cellular Therapy Applications

Lara J. Ausubel, Patricia M. Lopez, and Larry A. Couture

12. Culture of Human Pluripotent Stem Cells on Glass Slides for High-Resolution Imaging

Victoria Fox

Section IV. Characterization

13. Classical Cytogenetics: Karyotyping Techniques

Steven E. Bates

14. FISH Analysis of Human Pluripotent Stem Cells

Suzanne E. Peterson, Jerold Chun, and Jeanne Loring

15. Immunocytochemical Analysis of Human Pluripotent Stem Cells

Hubert E. Nethercott, David J. Brick, and Philip H. Schwartz

16. Flow Cytometric Analysis of Human Pluripotent Stem Cells

Mirabelle SH. Ho, Andrew Fryga, and Andrew L. Laslett

17. The Teratoma Assay: An In vivo Assessment of Pluripotency

Robin L. Wesselschmidt

18. Detection of Copy Number Variation using SNP Genotyping

Gregory M. Cooper and Heather C. Mefford

19. Genome-wide Epigenetic Analysis of Human Pluripotent Stem Cells by ChIP and ChIP-Seq

Michael J. Hitchler and Judd C. Rice

20. Basic Approaches to Gene Expression Analysis of Stem Cells by Microarrays

Bernhard Schuldt, Qiong Lin, Franz-Josef Müller, and Jeanne F. Loring

21. Development of High Content Screening Approaches and Analysis for Human Pluripotent Stem Cells

Sean P. Sherman, Jackelyn A. Alva, Kaushali Thakore-Shah, and April D. Pyle

22. Quantitative Proteome and Phosphoproteome Analysis of Human Pluripotent Stem Cells

Javier Muñoz and Albert J. R. Heck

Section V. Genetic Manipulation (Gene Modulation)

23. Lentivirus-mediated Modification of Pluripotent Stem Cells

Ruchi Bajpai

24. Nucleofection of Human Embryonic Stem Cells

Helen Fong, K. A. Hohenstein Elliott, Leslie F. Lock, and Peter J. Donovan

25. Non-viral Gene Delivery in Neural Progenitors Derived from Human Pluripotent Stem Cells

Sujoy K. Dhara, Anirban Majumder, Mahesh C. Dodla, and Steven L. Stice

26. Gene Targeting in Human Pluripotent Stem Cells

Ying Liu and Mahendra Rao

27. Episomal Transgene Expression in Pluripotent Stem Cells

Michele M.P. Lufino, Anna R. Popplestone, Sally A. Cowley, Pauline A.H. Edser, William S. James, and Richard Wade-Martins

Section VI: Differentiation

28. The Generation of Embryoid Bodies from Feeder-based or Feeder-free Human Pluripotent Stem Cell Cultures

Alexander E. Stover and Philip H. Schwartz

29. Derivation of Oligodendrocyte Progenitor C