Methods Development
Table of Contents
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. Immunocyhemical 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 Cells from Human Embryonic Stem Cells
Jason Sharp, Maya Hatch, Gabriel Nistor, and Hans Keirstead
30. Directed Differentiation of Dopamine Neurons From Human Pluripotent Stem Cells
Lixiang Ma, Yan Liu, and Su-Chun Zhang
31. Methods for the Derivation and Use of Cardiomyocytes from Human Pluripotent Stem Cells
Wei-Zhong Zhu, Benjamin Van Biber, and Michael A. Laflamme
32. In Vivo Evaluation of Putative Hematopoietic Stem Cells Derived from Human Pluripotent Stem Cells
Melinda K. Hexum, Xinghui Tian, and Dan S. Kaufman
33. Differentiation of Dendritic Cells from Human Embryonic Stem Cells
Kathryn M. Silk, Su-Yi Tseng, Kevin P. Nishimoto, Jane S. Lebkowski, Anita Reddy, and Paul J. Fairchild
Other Works by Dr. Schwartz in Methods Development
Table of Contents
Foreword by James Battey
Foreword by Jeannie Fontana
Preface
Part I: Basic methods: Maintenance of stem cell cultures
1.Human Embryonic Stem Cell Culture
2.Feeder layers and feeder-free culture
3.Mouse Embryonic Fibroblast Feeder Cells
4.Cryopreservation of hESC
Part II: Basic methods: Characterization of stem cells
5.Classical Cytogenetics: Karyotyping
6.Spectral Karyotyping and Fluorescent in situ Hybridization
7.Using SNP Genotyping to Identify Cell Lines, Determine Genomic Abnormalities and Monitor DNA Methylation Patterns
8.FACS/Cell sorting
9.Fluorescence Immunocytochemical Analysis of Stem Cells
10.Characterization of Stem Cells Using RT-PCR
11.Gene Expression Profiling of Stem Cells by Microarray
12.Teratomas produced from Human Embryonic Stem Cells Xenografted into Immunodeficient Mice
13.Generation of HESC-Derived Teratomas
Part III: Differentiation of stem cells
14.“Embryoid Body” Formation and Neuroepithelia Differentiation
15.Motor Neuron and Dopamine Neuron Differentiation
16.Oligodendrocyte Differentiation from hESC
17.Cardiac
18.Hematopoietic
Part IV: Genetic manipulation of stem cells
19.Genetic Manipulation of hESC: Lentivirus Vectors
20.Methods developed for mouse ES cells
Part V: Advanced methods
21.Derivation of human ES cells
22.Neural Stem Cell Culture
23.Transplantation of stem cells
24.In vitro fertilization
25.Functional characterization-neural cells
Part VI: Practical issues in establishing a human ES cell laboratory
26.Setting Up a Facility for hESC Research
Part VII: Essays on stem cell biology
27.Intellectual Property: Owning the Stem Cell
28.Ethical Concerns for Stem Cell Research
29.Guidelines for hESC Research Oversight (ESCRO) Committees