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This manual is a comprehensive compilation of "methods that work" for deriving, characterizing, and differentiating hPSCs, written by the researchers who developed and tested th… Read more
LIMITED OFFER
Immediately download your ebook while waiting for your print delivery. No promo code needed.
This manual is a comprehensive compilation of "methods that work" for deriving, characterizing, and differentiating hPSCs, written by the researchers who developed and tested the methods and use them every day in their laboratories. The manual is much more than a collection of recipes; it is intended to spark the interest of scientists in areas of stem cell biology that they may not have considered to be important to their work. The second edition of the Human Stem Cell Manual is an extraordinary laboratory guide for both experienced stem cell researchers and those just beginning to use stem cells in their work.
Foreword 1
Foreword 2
Preface 1
Preface 2
List of Contributors
PART 1: Basic Methods
Chapter 1. Culturing Human Pluripotent Stem Cells on a Feeder Layer
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Recipes
Reading List
Chapter 2. Preparation of Mouse Embryonic Fibroblast Feeder Cells
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Recipes
Reading List
Chapter 3. Culture of Human Pluripotent Stem Cells in Feeder-Free Conditions
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Recipes
Quality Control Methods
Reading List
Chapter 4. Contamination
Editor’s Commentary
Overview
Identifying Contamination
Controlling Contamination
Reagents and Supplies
Reading List
Chapter 5. Cell Culture Medium
Editor’s Commentary
Overview
Design of Culture Medium
Commercially Available Stem Cell Media
Assessing Medium Efficacy
Perspective
Reading List
Chapter 6. Cryopreservation of Human Embryonic Stem Cells
Editor’s Commentary
Overview
Procedures
Pitfalls and Advice
Alternative Procedures
Equipment
Reagents and Supplies
Recipes
Reading List
Chapter 7. Setting up a Research Scale Laboratory for Human Pluripotent Stem Cells
Editor’s Commentary
Overview
Laboratory
Record Keeping
Equipment Maintenance
Safety
Quality Control
Cell Banking
Reading List
Chapter 8. Biobanks for Pluripotent Stem Cells
Editor’s Commentary
Overview
Banking of Human Pluripotent Stem Cells
Stem Cell Providers
Future of Banking and New iPSC Initiatives
Reading List
PART 2: Induced Pluripotent Stem Cell Technology
Chapter 9. Isolation of Human Dermal Fibroblasts from Biopsies
Editor’s Commentary
Overview
Procedure
Alternative Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Recipes
Reading List
Chapter 10. Human Induced Pluripotent Stem Cell Generation
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Recipes
Quality Control Methods
Acknowledgments
Reading List
Chapter 11. Integration-Free Method for the Generation of Human Induced Pluripotent Stem Cells
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Recipes
Acknowledgments
Reading List
Chapter 12. Methods for Evaluating Human Induced Pluripotent Stem Cells
Editor’s Commentary
Overview
Procedures
Equipment
Reagents and Supplies
Acknowledgments
Reading List
PART 3: Characterization of Human Pluripotent Stem Cells
Chapter 13. Classical Cytogenetics
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Recipes
Acknowledgments
Reading List
Chapter 14. SNP Genotyping to Detect Genomic Alterations in Human Pluripotent Stem Cells
Editor’s Commentary
Overview
Procedures
Alternative procedures
Pitfalls and Advice
Equipment and Supplies
Quality Control Methods
Reading List
Chapter 15. Analysis and Purification Techniques for Human Pluripotent Stem Cells
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Reading list
Chapter 16. Immunocytochemical Analysis of Human Stem Cells
Editor’s Commentary
Overview
Procedures
Pitfalls and Advice
Digital Images of Fluorescent Cells
Equipment
Reagents and Supplies
Recipes
Reading List
Chapter 17. Analysis of Genome-Wide Gene Expression Data from Microarrays and Sequencing
Editor’s Commentary
Overview
Experimental Design
Critical Statistical Concepts
Microarray Data Analysis
RNA Sequencing (RNA-Seq)
Pitfalls and Advice
Reading List
Chapter 18. PluriTest Molecular Diagnostic Assay for Pluripotency in Human Stem Cells
Editor’s Commentary
Overview
Procedures
Pluritest Output
Novelty Score
Density Plot
Quality Control Outputs
Data, Experimental Design, and Formats
Pitfalls and Advice: Frequently Asked Questions
Reading List
Chapter 19. Epigenetics
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Equipment
Reagents and Supplies
Recipes
ChIP Antibodies
Quality Control Methods
Reading List
Chapter 20. Epigenetics
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Equipment, Reagents, and Supplies
Quality Control Methods
Reading List
Chapter 21. Generation of Human Pluripotent Stem Cell-Derived Teratomas
Editor’s Commentary
Overview
Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Reading List
Chapter 22. Characterization of Human Pluripotent Stem Cell-Derived Teratomas
Editor’s Commentary
Overview
Procedures
hiPSC-derived Teratomas
Reading List
PART 4: Differentiation
Chapter 23. Spontaneous Differentiation of Human Pluripotent Stem Cells via Embryoid Body Formation
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Recipes
Reading List
Chapter 24. Differentiation of Human Pluripotent Stem Cells into Neural Progenitors
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Reagents and Supplies
Recipes
Reading List
Chapter 25. Dopaminergic Neuronal Differentiation of Human Pluripotent Stem Cells
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Recipes
Reading List
Chapter 26. Directed Differentiation of Human Pluripotent Stem Cells to Oligodendrocyte Progenitor Cells
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Reagents and Supplies
Recipes
Quality Control Methods
Reading List
Chapter 27. Cardiomyocyte Differentiation of Human Pluripotent Stem Cells
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Recipes
Quality Control Methods
Reading List
Chapter 28. Directed Differentiation of Human Pluripotent Stem Cells into Fetal-Like Hepatocytes
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Recipes
Quality Control Methods
Reading List
PART 5: Genetic Manipulation
Chapter 29. Transfection of Human Pluripotent Stem Cells by Electroporation
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Recipes
Quality Control Methods
Reading List
Chapter 30. Lentiviral Vector Systems for Transgene Delivery
Editor’s Commentary
Overview
Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Recipes
Reading List
Chapter 31. Gene Targeting by Homologous Recombination in Human Pluripotent Stem Cells
Editor’s Commentary
Overview
Procedures
Reagents and Supplies
Recipes
Reading List
Chapter 32. Genetic Manipulation of Human Pluripotent Stem Cells Using Zinc Finger Nucleases
Editor’s Commentary
Overview
Procedures
Pitfalls and Advice
Reagents and Supplies
Recipes
Reading List
PART 6: Stem Cell Transplantation
Chapter 33. Intraspinal Transplantation of Human Neural Stem Cells
Editor’s Commentary
Overview
Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Recipes
Quality Control Methods
Reading List
Chapter 34. Transplanting Stem Cells into the Brain
Editor’s Commentary
Overview
Procedures
Pitfalls and Caveats
Equipment
Reagents and Supplies
Recipes
Quality Control Methods
Reading List
PART 7: Derivation of Human Pluripotent Stem Cells
Chapter 35. Development of Human Blastocysts
Editor’s Commentary
Overview
Laws and Ethics
Background
Procedures
Equipment
Reagents and Supplies
Recipes
Reading List
Chapter 36. Derivation of Human Embryonic Stem Cells from Blastocysts
Editor’s Commentary
Overview
Procedures
Alternative Procedures
Pitfalls and Advice
Equipment
Reagents and Supplies
Recipes
Quality Control Methods
Reading List
PART 8: Stem Cells and Society
Chapter 37. Stem Cell Patents
Editor’s Commentary
Overview
What is a Patent?
Obtaining a Patent on an Invention
The America Invents Act of 2011
Patent Licensing Policies
Material Transfer
Patent Pools
Stem Cell Patents
Human Embryonic Stem Cell Patents
Induced Pluripotent Stem Cell Patents
Reading List
Chapter 38. Ethics of Human Pluripotent Stem Cell Research and Development
Editor’s Commentary
Overview
Ethical Issues for Human Pluripotent Stem Cell Research
Informed Consent
Early Clinical Trials and Stem Cell Tourism
Review Committees
Scientific Responsibilities
Reading List
Index
SP
JL