Genetic Modification of Hematopoietic Stem Cells

1. Front Matter

   Pages i-xxi

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2. Immunomagnetic Enrichment of Human and Mouse Hematopoietic Stem Cells for Gene Therapy Applications

   • Guillermo Guenechea, Jose C. Segovia, Juan A. Bueren

   Pages 1-11

3. Isolation of Human and Mouse Hematopoietic Stem Cells

   • Yuk Yin Ng, Miranda R.M. Baert, Edwin F.E. de Haas, Karin Pike-Overzet, Frank J.T. Staal

   Pages 13-21

4. Murine Hematopoietic Stem Cell Transduction Using Retroviral Vectors

   • Ute Modlich, Axel Schambach, Zhixiong Li, Bernhard Schiedlmeier

   Pages 23-31

5. Genetic Modification of Human Hematopoietic Cells: Preclinical Optimization of Oncoretroviral-mediated Gene Transfer for Clinical Trials

   • Tulin Budak-Alpdogan, Isabelle Rivière

   Pages 33-58

6. Short-Term Culture of Human CD34+ Cells for Lentiviral Gene Transfer

   • Francesca Santoni de Sio, Luigi Naldini

   Pages 59-70

7. T Cell Culture for Gammaretroviral Transfer

   • Sebastian Newrzela, Brandenburg Gunda, Dorothee von Laer

   Pages 71-82

8. Retroviral Transduction of Murine Primary T Lymphocytes

   • James Lee, Michel Sadelain, Renier Brentjens

   Pages 83-96

9. Lentiviral Vector Gene Transfer into Human T Cells

   • Els Verhoeyen, Caroline Costa, Francois-Loic Cosset

   Pages 97-114

10. DNA Transposons for Modification of Human Primary T Lymphocytes

    • Xin Huang, Andrew Wilber, R. Scott McIvor, Xianzheng Zhou

    Pages 115-126

11. Retroviral Gene Transfer into Primary Human Natural Killer Cells

    • Evren Alici, Tolga Sutlu, M. Sirac Dilber

    Pages 127-137

12. Lentiviral Vector-Mediated Genetic Programming of Mouse and Human Dendritic Cells

    • Renata Stripecke

    Pages 139-158

13. In Situ (In Vivo) Gene Transfer into Murine Bone Marrow Stem Cells

    • Dao Pan

    Pages 159-169

14. In Vivo and Ex Vivo Gene Transfer in Thymocytes and Thymocyte Precursors

    • Oumeya Adjali, Amélie Montel-Hagen, Louise Swainson, Sophie Marty, Rita Vicente, Cedric Mongellaz et al.

    Pages 171-190

15. Design and Production of Retro- and Lentiviral Vectors for Gene Expression in Hematopoietic Cells

    • Axel Schambach, William P. Swaney, Johannes C.M. van der Loo

    Pages 191-205

16. Knock-Down of Gene Expression in Hematopoietic Cells

    • Michaela Scherr, Letizia Venturini, Matthias Eder

    Pages 207-219

17. The Use of Retroviral Vectors for tet-Regulated Gene Expression in Cell Populations

    • Rainer Löw

    Pages 221-242

18. Detection of Replication Competent Retrovirus and Lentivirus

    • Lakshmi Sastry, Kenneth Cornetta

    Pages 243-263

19. Release Testing of Retroviral Vectors and Gene-Modified Cells

    • Diana Nordling, Anne Kaiser, Lilith Reeves

    Pages 265-279

20. Copy Number Determination of Genetically-Modified Hematopoietic Stem Cells

    • Todd Schuesler, Lilith Reeves, Christof von Kalle, Elke Grassman

    Pages 281-298

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).

• Complex nucleic acids
• DNA
• Gene medicine
• Gene transfer
• Hematology
• Hematopoiesis
• Microarray
• Mutagenesis
• PCR
• Proteomics
• Termination
• gene expression
• gene therapy
• genes
• hybridization

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)

• Department of Experimental Hematology, Hannover Medical School, Hannover, Germany

  Christopher Baum

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