Essential Stem Cell Methods

Author： Lanza, Robert;Klimanskaya, Irina

Publishers： Elsevier Science

Publishing year： 2011

ISBN： 9780080885254

Subject： [Q 生物科学]

Language： ENG

Introduction

This is a fast-moving field, and these detailed methods will help drive advances in stem cell research. The editors have hand selected step-by-step methods from researchers with extensive reputations and expertise.

This volume, as part of the Reliable Lab Solutions series, delivers busy researchers a handy, time-saving source for the best methods and protocols in stem cells.

* Provides powerful research opportunities for those interested in perusing work in pluripotent stem cells, disease modeling, and other aspects of basic stem cell research

* Refines, organizes and updates popular methods from flagship series, Methods in Enzymology

*Highlights top downloads, enhanced with author tips and tricks and pitfalls to avoid

Chapter

  • Front Cover

  • Essential Stem Cell Methods

  • Copyright Page

  • Contents

  • Contributors

  • Preface

  • Section I: Organ-Derived Stem Cells

  • Chapter 1: Neural Stem Cells: Isolation and Characterization

  • 1. Introduction

  • 2. Reagents and Instrumentation

  • 3. Methods

  • Acknowledgments

  • References

  • Chapter 2: Neural Stem Cells and Their Manipulation

  • 1. Introduction

  • 2. Adult Niches for Stem Cells In Vivo

  • 3. In Vitro Manipulation of NSCs

  • 4. Conclusions and Projections

  • 5. Sample Protocols for the Culture and Characterization of NSCs

  • 6. Culture of Rodent/Murine NSCs

  • 7. Culture of Human NSCs

  • 8. ICC of Markers to Identify Stem Cells and Differentiation Products

  • Acknowledgments

  • References

  • Chapter 3: Retinal Stem Cells

  • 1. Introduction

  • 2. Materials and Methods

  • 3. Overview

  • References

  • Chapter 4: Dental Pulp Stem Cells

  • 1. Introduction

  • 2. Identification of Dental Pulp Stem Cells (DPSCs)

  • 3. Isolation of DPSCs

  • 4. Differentiation of DPSCs

  • References

  • Chapter 5: Mouse Spermatogonial Stem Cells: Culture and Transplantation

  • 1. Introduction

  • 2. Spermatogonial Stem Cells (SSCs)

  • 3. SSC Transplantation

  • 4. SSC Culture

  • 5. siRNA Transfection of Cultured Mouse SSCs

  • 6. Implications

  • Acknowledgments

  • References

  • Chapter 6: Stem Cells in the Adult Lung

  • 1. Introduction

  • 2. Anatomical and Cellular Diversity of the Adult Lung

  • 3. Stem Cell Phenotypes and Niches in the Adult Lung

  • 4. In Vivo Injury Models of the Lung

  • 5. Ex Vivo Epithelial Tracheal Xenograft Model to Study Stem Cells Expansion in the Proximal Airway

  • 6. In Vitro Colony-Forming Efficiency (CFE) Assay to Characterize Stem/progenitor Cell Populations in Conducting Airway Epithel

  • 7. Models to Study Stem/Progenitor Cells of Airway SMG

  • Acknowledgments

  • References

  • Chapter 7: Pancreatic Cells and Their Progenitors

  • 1. Introduction

  • 2. Pancreas Development

  • 3. Origins of Beta Cells During Postnatal Life

  • 4. Preexisting Beta Cells

  • 5. Ducts

  • 6. Acini

  • 7. Bone Marrow Cells

  • 8. Adult Stem Cells

  • 9. Dedifferentiation of Beta Cells

  • 10. Summary and Perspective

  • 11. Methods: Design of a Lineage Tracing Experiment in Mice

  • References

  • Chapter 8: Pluripotent Stem Cells from Germ Cells: Derivation and Maintenance

  • 1. Introduction

  • 2. Germ Cell Development

  • 3. EGC Derivation

  • 4. Characterization of EG Cultures

  • 5. EB Formation and Analysis

  • 6. EB-Derived Cell Formation

  • Acknowledgments

  • References

  • Chapter 9: Pluripotent Stem Cells from Amniotic Fluid and Placenta

  • 1. Introduction

  • 2. Amniotic Fluid and Placenta in Developmental Biology

  • 3. Amniotic and Placental Cells for Therapy-A New Connection

  • 4. Isolation and Characterization of Progenitor Cells

  • 5. Differentiation of Amniotic and Placental-Derived Progenitor Cells

  • 6. Conclusion

  • References

  • Chapter 10: Hematopoietic Stem and Progenitor Cells from Cord Blood

  • 1. Introduction: Cord Blood Transplantation

  • 2. Methodologies for Assessing Hematopoietic Progenitor and Stem Cells, as Well as EPC, Present in Human Cord Blood

  • 3. Uses of Above Assays

  • References

  • Chapter 11: Hematopoietic Stem Cells from Bone Marrow: Purification and Functional Analysis

  • 1. Introduction

  • 2. Other HSC Surface Markers: Tie-2, Endoglin, and the SLAM Family Receptors

  • 3. Fluorescent Dye Efflux in HSCs and the SPKLS

  • 4. Characteristics of HSCs from Different Purification Schemes

  • 5. Protocol of HSC Sorting with Hoechst 33342 Staining (SP Population)

  • References

  • Chapter 12: Microarray Analysis of Stem Cells and Their Differentiation

  • 1. Introduction

  • 2. Overview of Microarray Technology

  • 3. Experimental Design

  • 4. Confirmation Studies

  • 5. Examples of Microarray Experiments for Stem Cell Biology and Differentiation

  • 6. Identification of "Stemness"

  • 7. Differentiation

  • 8. Stem Cell Niches

  • 9. Future Directions

  • References

  • Chapter 13: Tissue Engineering Using Adult Stem Cells

  • 1. Introduction

  • 2. Biomaterials

  • 3. Angiogenic Factors

  • 4. Adult Stem Cells for Tissue Engineering

  • 5. Conclusion

  • References

  • Chapter 14: Tissue Engineering Using Mesenchymal Stem Cells

  • 1. MSCs: Definition and Therapeutic Promise

  • 2. Isolation and Expansion of MSCs

  • 3. Multilineage Differentiation of MSCs

  • 4. Clinical Translation of MSC-Based Therapies

  • 5. Conclusions

  • Acknowledgments

  • References

  • Section II: Embryonic Stem Cells and Their Derivatives

  • Chapter 15: Mouse Embryonic Stem Cells

  • 1. Historical Overview

  • 2. Factors Affecting the Effi;ciency of Mouse ES Cell Establishment

  • 3. Factors Affecting the Contribution of Mouse ES Cell to Chimeric Embryos

  • 4. Critical Events During Mouse ES Cell Establishment

  • 5. Freezing of ES Cell Lines

  • 6. Characterization

  • 7. Protocols

  • Acknowledgment

  • References

  • Chapter 16: Human Embryo Culture

  • 1. Introduction

  • 2. Human Embryo Development

  • 3. Embryo Biopsy

  • 4. Human Embryo Culture

  • 5. Results

  • References

  • Chapter 17: Human Embryonic Stem Cells: Derivation and Maintenance

  • 1. Introduction

  • 2. Derivation of hES Cell Lines

  • 3. Maintenance of hES Cells

  • 4. Conclusion

  • Acknowledgments

  • References

  • Chapter 18: Human Embryonic Stem Cells: Characterization and Evaluation

  • 1. Introduction

  • 2. Characterization of Undifferentiated hESCs

  • 3. Conclusion

  • Acknowledgments

  • References

  • Chapter 19: Human Embryonic Stem Cells: Feeder-Free Culture

  • 1. Introduction

  • 2. Methods for Feeder Layer-Free Culture of hESCs

  • Acknowledgments

  • References

  • Chapter 20: Neural Stem Cells, Neurons, and Glia from Embryonic Stem Cells

  • 1. Introduction

  • 2. Protocols

  • 3. Summary

  • 4. Media and Reagents

  • Acknowledgments

  • References

  • Chapter 21: Hematopoietic Cell Differentiation from Embryonic Stem Cells

  • 1. Introduction

  • 2. Methods

  • References

  • Chapter 22: Cardiomyocyte Differentiation from Embryonic Stem Cells

  • 1. Introduction

  • 2. Embryoid Body Generation

  • 3. Cardiomyocyte Differentiation

  • 4. Enrichment of Cardiomyocytes

  • 5. Conclusion

  • References

  • Chapter 23: Insulin-Producing Cells from Mouse Embryonic Stem Cells

  • 1. Introduction

  • 2. Materials and Methods

  • 3. Results

  • 4. Summary

  • References

  • Chapter 24: Transgene Expression and RNA Interference in Embryonic Stem Cells

  • 1. Retrovirus Expression Vectors and ESCs

  • 2. RNA Interference and ESCs

  • 3. siRNA Expression Vector Design

  • 4. Retrovirus Production

  • 5. Retroviral and Lentiviral Gene Transfer into Mouse and Human ESCs

  • 6. Transgene and siRNA Expression in Mouse and hESCs

  • 7. Biotechnological and Medical Applications

  • Acknowledgments

  • References

  • Chapter 25: Lentiviral Vector-Mediated Gene Delivery into Human Embryonic Stem Cells

  • 1. Introduction

  • 2. Design of HIV-1 Based Vectors for Transduction of hESCs

  • 3. Generation of Recombinant Viral Particles

  • 4. Transduction of hESCs

  • 5. Measurement of Transduction Efficiency

  • 6. Enrichment for Transduced hESCs Expressing High Levels of the Transgene

  • 7. Determination of Viral Titer

  • References

  • Chapter 26: Engineering Embryonic Stem Cells with Recombinase Systems

  • 1. Introduction

  • 2. Site-Specific Recombination

  • 3. Designing Substrates for Site-Specific Recombination

  • 4. Generation of Conditional Alleles

  • 5. Recombinase Mediated Cassette Exchange (RMCE)

  • 6. Molecular Switches

  • 7. Protocols

  • Acknowledgments

  • References

  • Chapter 27: Tissue Engineering Using Embryonic Stem Cells

  • 1. Introduction

  • 2. Special Considerations when using hESCs as the Cell Source for TE

  • 3. Growing hESCs in Defined Animal-Free Conditions

  • 4. Obtaining the Desired Cell Population

  • 5. Choosing the Right Scaffold

  • 6. Scaling-Up a Regulable Bioprocess

  • 7. hESC-Derived Connective Tissue Progenitors for TE

  • 8. Culture and Maintenance of hESC on MEF Feeders

  • 9. Harvesting Samples for Analyses

  • References

  • Index

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