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Genetic Modification of Hematopoietic Stem Cells

  • Kartonierter Einband
  • 524 Seiten
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With the huge potential of gene transfer into hematopoietic stem cells, active research in this field has become critically import... Weiterlesen
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With the huge potential of gene transfer into hematopoietic stem cells, active research in this field has become critically important. In this volume, leading scientists in the field provide a compendium of protocols which cover the subject comprehensively.

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).

Provides an easily accessible reference volume for genetic modification of hematopoietic cells using various state-of-the-art vector systems

Comprehensive guide on functional, phenotypic and genotypic screening of gene-modified cells written by leading researchers in the field

Contains helpful insights into preclinical data acquisition and documentation for gene therapy, including regulatory comments from the EU and US perspectives

Covers a wide range of cell-based and animal assay formats and systems


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™ 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.

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 Rivière 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, Amélie Montel-Hagen, Louise Swainson, Sophie Marty, Rita Vicente, Cedric Mongellaz, Chantal Jacquet, Valérie 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 Löw 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


Titel: Genetic Modification of Hematopoietic Stem Cells
Untertitel: Methods and Protocols
EAN: 9781617378669
ISBN: 1617378666
Format: Kartonierter Einband
Herausgeber: Humana Press
Anzahl Seiten: 524
Gewicht: 1267g
Größe: H280mm x B210mm x T28mm
Jahr: 2011
Auflage: Softcover reprint of hardcover 1st ed. 2009

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