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Stem Cell Tools and Other Experimental Protocols
1st Edition, Volume 420 - December 12, 2006
Editors: Robert Lanza, Irina Klimanskaya
Language: English
Hardback ISBN:9780123736512
9 7 8 - 0 - 1 2 - 3 7 3 6 5 1 - 2
eBook ISBN:9780080469645
9 7 8 - 0 - 0 8 - 0 4 6 9 6 4 - 5
This is the third of three planned volumes in the Methods in Enzymology series on the topic of stem cells. This volume is a unique anthology of stem cell techniques written by exp…Read more
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This is the third of three planned volumes in the Methods in Enzymology series on the topic of stem cells. This volume is a unique anthology of stem cell techniques written by experts from the top laboratories in the world. The contributors not only have hands-on experience in the field but often have developed the original approaches that they share with great attention to detail. The chapters provide a brief review of each field followed by a “cookbook” and handy illustrations. The collection of protocols includes the isolation and maintenance of stem cells from various species using “conventional” and novel methods, such as derivation of ES cells from single blastomeres, differentiation of stem cells into specific tissue types, isolation and maintenance of somatic stem cells, stem cell-specific techniques and approaches to tissue engineering using stem cell derivatives. The reader will find that some of the topics are covered by more than one group of authors and complement each other. Comprehensive step-by-step protocols and informative illustrations can be easily followed by even the least experienced researchers in the field, and allow the setup and troubleshooting of these state-of-the-art technologies in other laboratories.
Provides complete coverage spanning from derivation/isolation of stem cells, and including differentiation protocols, characterization and maintenance of derivatives and tissue engineering
Presents the latest most innovative technologies
Addresses therapeutic relevance including FDA compliance and tissue engineering
Biochemists, cell and molecular biologists, and bioengineers working with stem cells.
Dedication
Contributors to Volume 420
Preface
Foreword
Volumes in Series
Section I: In Vitro Experimentation and Research Tools
[1]: Human Embryo Culture
Abstract
Introduction
Human Embryo Development
Human Embryo Culture: General Considerations
Protocols
Results
Acknowledgments
[2]: Characterization and Evaluation of Human Embryonic Stem Cells
Abstract
Introduction
Characterization of Undifferentiated hESCs
Conclusion
Acknowledgments
[3]: Feeder-Free Culture of Human Embryonic Stem Cells
Abstract
Introduction
Methods for Feeder-Free Culture of hESCs
Acknowledgments
[4]: Transgene Expression and RNA Interference in Embryonic Stem Cells
Abstract
Retrovirus Expression Vectors and Embryonic Stem Cells
RNA Interference and ESCs
siRNA Expression Vector Design
Retrovirus Production
Retroviral and Lentiviral Gene Transfer into Mouse and Human ESCs
Transgene and siRNA Expression in Mouse and Human ESCs
Biotechnological and Medical Applications
Acknowledgments
[5]: Lentiviral Vector–Mediated Gene Delivery into Human Embryonic Stem Cells
Abstract
Introduction
Development of Lentiviral-Based Vectors
Design of HIV-1–Based Vectors
Transduction of hESCs by Lentiviral-Based Vectors
Potential Applications of Gene Delivery into hESCs by Lentiviral-Based Vectors
Design of HIV-1–Based Vectors for Transduction of hESCs
[6]: The Use of Retroviral Vectors for Gene Transfer into Hematopoietic Stem Cells
Publisher Summary
Introduction
Viral Pseudotype
Vector Design
Packaging Cell Lines
Clinical Applications
Generation of Retrovirus Packaging Cell Lines
Summary and Future Directions
Acknowledgments
[7]: Engineering Embryonic Stem Cells with Recombinase Systems
Abstract
Introduction
Site-Specific Recombination
Large Serine Recombinases
Designing Substrates for Site-Specific Recombination
Generation of Conditional Alleles
Recombinase-Mediated Cassette Exchange (RMCE)
Molecular Switches
Protocols
[8]: Gene Trapping in Embryonic Stem Cells
Abstract
Introduction
Common Types of Gene Trap Vectors in IGTC
A User’s Guide to the International Gene Trap Consortium Resource
Investigator-Initiated Screens
[9]: GeneChips in Stem Cell Research
Abstract
Introduction
Protocol
ESC Differentiation
ESC Genes and In Vivo Regeneration
Cell Screening
Integration and Meta-Analysis of GeneChip and Microarray Data
Applications to Cancer
Conclusion
Summary
Acknowledgments
Appendix 1 Sample Analysis
[10]: Microarray Analysis of Stem Cells and Differentiation
Abstract
Introduction
Overview of Microarray Technology
Total RNA Isolation and Clean-Up
RNA Amplification
Direct Labeling of RNA for Microarray Hybridization
Indirect Labeling of RNA for Microarray Hybridization
Examples of Microarray Experiments for Stem Cell Biology and Differentiation
Identification of Stemness
Differentiation
Stem Cell Niches
Future Directions
[11]: Purification of Hematopoietic Stem Cells Using the Side Population
Abstract
Introduction
Protocol of HSC Sorting with Hoechst 33342 Staining (SP)
[12]: Cellular Reprogramming
Abstract
Introduction
Streptolysin O–Mediated Cell Permeabilization and Uptake of Reprogramming Cell Extracts
Cell Permeabilization and the Reprogramming Reaction
Preparation of Reprogramming Cell Extracts
Acknowledgments
Section II. Tissue Engineering and Regenerative Medicine
[13]: Tissue Engineering Using Adult Stem Cells
Abstract
Introduction
Biomaterials
Angiogenic Factors
Adult Stem Cells for Tissue Engineering
Conclusion
[14]: Tissue Engineering Using Human Embryonic Stem Cells
Abstract
Introduction
Special Considerations When Using hESCs as the Cell Source for TE
Growing hESCs in Defined Animal-Free Conditions
Obtaining the Desired Cell Population
Choosing the Right Scaffold
Scaling Up a Regulatable Bioprocess
hESC-Derived Connective Tissue Progenitors for TE
Preparation of MEF Feeder Layers
Starting hESC Culture
Passaging hESCs
Choosing the Right Scaffold for Connective Tissue Engineering
Harvesting Samples for Analyses
Electron Microscopy
Histological Analysis
[15]: Engineering Cardiac Tissue from Embryonic Stem Cells
Abstract
Introduction
Liquid Collagen–Based Cardiac Tissue Engineering
Conclusions
[16]: Mesenchymal Stem Cells and Tissue Engineering
Abstract
MSCs: Definition and Therapeutic Promise
Isolation and Expansion of MSCs
Multilineage Differentiation of MSCS
Clinical Translation of MSC-Based Therapies
Conclusions
Acknowledgments
[17]: Bone Reconstruction with Bone Marrow Stromal Cells
Abstract
Introduction
Cell Source for Bone Engineering
Animal Models for BMSC-Mediated Bone Engineering
Clinical Application of Engineered Bone
Protocol for Dog Cranial Bone Engineering
Protocol for Goat Femoral Bone Engineering (Zhu et al., 2006)
Acknowledgment
[18]: Engineering Three-Dimensional Tissue Structures Using Stem Cells
Abstract
Introduction
Protocols for Differentiating ESCs
Protocol for Scaffold Seeding
[19]: Immunogenicity of Embryonic Stem Cells and Their Progeny
Abstract
Introduction
Generation of Trimera Mice
Transplantation of hESCs and Derivatives into Trimera Mice
Assessing the Immune Response against Engrafted hESCs
Results and Discussion
Acknowledgments
[20]: Manufacturing Considerations for Clinical Uses of Therapies Derived from Stem Cells
Abstract
Introduction
U.S. Regulatory Framework for Therapies Derived from Stem Cells
The Need for a Science-Based Approach for the Product and the Process
Ancillary Materials
Residual Ancillary Materials in Final Product
Genetic Modification
Combination Products
Developing a Manufacturing Process That Is Consistent and Scalable
Aseptic Processing
Characterization
In-Process and Release Testing
Shipment
Handling of Cellular Product at Clinical Site
The Need for a Quality Program
Summary
Author Index
Subject Index
No. of pages: 520
Language: English
Edition: 1
Volume: 420
Published: December 12, 2006
Imprint: Academic Press
Hardback ISBN: 9780123736512
eBook ISBN: 9780080469645
RL
Robert Lanza
Robert Lanza is an American scientist and author whose research spans the range of natural science, from biology to theoretical physics. TIME magazine recognized him as one of the “100 Most Influential People in the World,” and Prospect magazine named him one of the Top 50 “World Thinkers.” He has hundreds of scientific publications and over 30 books, including definitive references in the fields of stem cells, tissue engineering, and regenerative medicine. He’s a former Fulbright Scholar and studied with polio-pioneer Jonas Salk and Nobel laureates Gerald Edelman (known for his work on the biological basis of consciousness) and Rodney Porter. He also worked closely (and co-authored papers in Science on self-awareness and symbolic communication) with noted Harvard psychologist BF Skinner. Dr. Lanza was part of the team that cloned the world’s first human embryo, the first endangered species, and published the first-ever reports of pluripotent stem cell use in humans.
Affiliations and expertise
Astellas Institute for Regenerative Medicine, Westborough, MA, USA
IK
Irina Klimanskaya
Affiliations and expertise
Advance Cell Technology, Worcester, MA, USA
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