Genetic Modification of Hematopoietic Stem Cells: Genetic Modification of Hematopoietic Stem Cells Preliminary Entry 2072
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Genetic Modification of Hematopoietic Stem Cells: Genetic Modification of Hematopoietic Stem Cells Preliminary Entry 2072 : Methods and Protocols

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Gene Transfer into Hematopoietic Cells: From Basic Science to Clinical Application Christopher Baum 1. The Potential of Gene Transfer into Hematopoietic Vectors with the potential for stable transgene integration are Cells widely used in basic hematology and clinical trials of gene me- cine. In basic research, both gain-of-function and loss-of-fu- tion situations of individual genes can be created by gene transfer, leading to a wide range of applications in developmental biology, stem cell biology, immunology, leukemia research, and human genetics. With the first evidence of successful modification of murine hematopoietic cells using retroviral gene vectors (1, 2), researchers have also explored the therapeutic potential of this approach. To date, the emerging discipline of gene therapy is a highly diversified field that offers entirely novel approaches to treat a great variety of human diseases (3). All hematopoietic cell types are of major interest in this context, since the modification of the hematopoietic stem cell population may potentially give rise to a completely transgenic hematopoiesis with the potential to cure genetic disorders or fight severe chronic infections, and the targeting of mature cells such as lymphocytes or antigen-p- senting dendritic cells offers all types of transient and semiper- nent modifications of the immune system. The unifying principle of gene medicine is the need to transfer complex nucleic acids cells that do not contribute to the germline (somatic cells).
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Product details

  • Hardback | 490 pages
  • 194 x 262 x 28mm | 1,179.33g
  • Totowa, NJ, United States
  • English
  • 2009 ed.
  • 30 Tables, black and white; 10 Illustrations, color; 74 Illustrations, black and white; XXII, 490 p. 84 illus., 10 illus. in color.
  • 1588299805
  • 9781588299802

Back cover copy

With the incredible potential of gene transfer into hematopoietic stem cells, active research in this field has become critically important. In Genetic Modification of Hematopoietic Stem Cells: Methods and Protocols, leading scientists in the field provide a compendium of protocols which cover the subject comprehensively, from the purification and culture of various types of hematopoietic cells for subsequent genetic modification by vector development and technical issues of small and large scale vector production, to the complex issue of monitoring and biosafety studies related to gene-modified hematopoiesis. Written in the highly successful Methods in Molecular Biology(TM) series format, the chapters in this volume present brief introductions to the topic, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and Notes sections, which allow the experts to highlight tips on troubleshooting and avoiding known pitfalls.



Unique and cutting-edge, Genetic Modification of Hematopoietic Stem Cells: Methods and Protocols is an ideal, thorough resource to promote further research and the implementation of investigator-driven clinical studies using gene-modified hematopoietic cells.
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Table of contents

1. Immunomagnetic Enrichment of Human and Mouse Hematopoietic Stem Cells for Gene Therapy Applications
Guillermo Guenechea, Jose C. Segovia, and Juan A. Bueren

2. Isolation of Human and Mouse Hematopoietic Stem Cells
Yuk Yin Ng, Miranda R.M. Baert, Edwin F.E. de Haas, Karin Pike-Overzet, and Frank J.T. Staal

3. Murine Hematopoietic Stem Cell Transduction Using Retroviral Vectors
Ute Modlich, Axel Schambach, Zhixiong Li, and Bernhard Schiedlmeier

4. Genetic Modification of Human Hematopoietic Cells: Preclinical Optimization of Oncoretroviral-Mediated Gene Transfer for Clinical Trials
Tulin Budak-Alpdogan and Isabelle Riviere

5. Short-Term Culture of Human CD34+ Cells for Lentiviral Gene Transfer
Francesca Santoni Di Sio and Luigi Nalidini

6. T Cell Culture for Gammaretroviral Transfer
Sebastian Newrzela, Gunda Brandenburg, and Dorothee von Laer

7. Retroviral Transduction of Murine Primary T Lymphocytes
James Lee, Michel Sadelain, and Renier Brentjens

8. Lentiviral Vector Gene Transfer into Human T Cells
Els Verhoeyen, Caroline Costa, and Francois-Loic Cosset

9. DNA Transposons for Modification of Human Primary T Lymphocytes
Xin Huang, Andrew C. Wilber, R. Scott McIvor, and Xianzheng Zhou

10. Retroviral Gene Transfer into Primary Human Natural Killer Cells
Evren Alici, Tolga Sutlu, and M. Sirac Dilber

11. Lentiviral Vector-Mediated Genetic Programming of Mouse and Human Dendritic Cells
Renata Stripecke

12. In Situ (In Vivo) Gene Transfer into Murine Bone Marrow Stem Cells
Dao Pan

13. In-vivo and ex-vivo Gene Transfer in Thymocytes and Thymocyte-Precursors
Oumeya Adjali, Amelie Montel-Hagen, Louise Swainson, Sophie Marty, Rita Vicente, Cedric Mongellaz, Chantal Jacquet, Valerie Zimmermann, andNaomi Taylor

14. Design and Production of Retro- and Lentiviral Vectors for Gene Expression in Hematopoietic Cells
Axel Schambach, William S. Swaney, and Johannes C. M. van der Loo

15. Knock-Down of Gene Expression in Hematopoietic Cells
Michaela Scherr, Letizia Venturini, and Matthias Eder

16. The Use of Retroviral Vectors for tet-Regulated Gene Expression in Cell Populations
Rainer Loew

17. Detection of Replication Competent Retrovirus and Lentivirus
Lakshmi Sastry and Kenneth Cornetta

18. Release Testing of Retroviral Vectors and Gene Modified Cells
Diana Nordling, Anne Kaiser, and Lilith Reeves

19. Copy Number Determination of Genetically Modified Hematopoietic Stem Cells
Todd Schuesler, Lilith Reeves, Christof von Kalle, and Elke Grassman

20. Tissue Procurement for Molecular Studies Using Laser-Assisted Microdissection
Ulrich Lehmann and Hans Kreipe

21. Leukemia Diagnosis in Murine Bone Marrow Transplantation Models
Zhixiong Li, Ute Modlich, and Anjali Mishra

22. Humanized Mouse Models to Study the Human Haematopoietic Stem Cell Compartment
Dominique Bonnet

23. Canine Models of Gene-Modified Hematopoiesis
Brian C. Beard and Hans-Peter Kiem

24. Detection of Retroviral Integration Sites by Linear Amplification Mediated PCR (LAM-PCR) and Tracking of Individual Integration Clones in Different Samples
Manfred Schmidt, Kerstin Schwarzwaelder, Cynthia Bartholomae, Hanno Glimm, and Christof von Kalle

25. Retroviral Insertion Site Analysis in Dominant Haematopoietic Clones
Olga S. Kustikova, Ute Modlich, and Boris Fehse

26. Tracking Gene-Modified T-Cells In vivo
Alessandra Recchia and Fulvio Mavilio

27. DNA Microarray Studies of Hematopoietic
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Review Text

From the reviews:

"This book provides a unique and comprehensive resource of protocols for the genetic modification of various hematopoietic cell types and up-to-date procedures for molecular and systemic monitoring. ... This book is a great help for the design of excellent research in basic hematology, oncology, genetics, and immunology, and also promote the implementation of investigator-driven clinical studies using gene-modified hematopoietic cells. The book represents an outstanding work, which should not be missed in all biomedical research laboratories dealing with gene therapy of hematopoietic cells." (C. Altaner, Neoplasma, June, 2009)

"This book provides a unique and comprehensive resource of protocols for the genetic modification of various hematopoietic cell types and up-to-date procedures for molecular and systemic monitoring. ... This book is a great help for the design of excellent research in basic hematology, oncology, genetics, and immunology, and also promote the implementation of investigator-driven clinical studies using gene-modified hematopoietic cells. The book represents an outstanding work, which should not be missed in all biomedical research laboratories dealing with gene therapy of hematopoietic cells." (C. Altaner, Neoplasma, April, 2009)

"Haematopoietic stem cells are attractive targets for gene therapy. ... Each chapter provides a brief review of the area, followed by a step-by-step guide to undertaking experiments and includes details of reagents required and a list of useful notes or comments of issues that may arise. Comprehensive reference citations provide sources of further information when required. The book should provide a road map for researchers, clinical investigators and regulators involved in modifying haematopoietic cells." (Waseem Qasim, Human Genetics, Vol. 128, May, 2010)
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Review quote

From the reviews:



"This book provides a unique and comprehensive resource of protocols for the genetic modification of various hematopoietic cell types and up-to-date procedures for molecular and systemic monitoring. ... This book is a great help for the design of excellent research in basic hematology, oncology, genetics, and immunology, and also promote the implementation of investigator-driven clinical studies using gene-modified hematopoietic cells. The book represents an outstanding work, which should not be missed in all biomedical research laboratories dealing with gene therapy of hematopoietic cells." (C. Altaner, Neoplasma, June, 2009)

"This book provides a unique and comprehensive resource of protocols for the genetic modification of various hematopoietic cell types and up-to-date procedures for molecular and systemic monitoring. ... This book is a great help for the design of excellent research in basic hematology, oncology, genetics, and immunology, and also promote the implementation of investigator-driven clinical studies using gene-modified hematopoietic cells. The book represents an outstanding work, which should not be missed in all biomedical research laboratories dealing with gene therapy of hematopoietic cells." (C. Altaner, Neoplasma, April, 2009)

"Haematopoietic stem cells are attractive targets for gene therapy. ... Each chapter provides a brief review of the area, followed by a step-by-step guide to undertaking experiments and includes details of reagents required and a list of useful notes or comments of issues that may arise. Comprehensive reference citations provide sources of further information when required. The book should provide a road map for researchers, clinical investigators and regulators involved in modifying haematopoietic cells."--- (Waseem Qasim, Human Genetics, Vol. 128, May, 2010)
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