Immunomagnetic Enrichment of Human and Mouse Hematopoietic Stem Cells for Gene Therapy Applications -- Isolation of Human and Mouse Hematopoietic Stem Cells -- Murine Hematopoietic Stem Cell Transduction Using Retroviral Vectors -- Genetic Modification of Human Hematopoietic Cells: Preclinical Optimization of Oncoretroviral-mediated Gene Transfer for Clinical Trials -- Short-Term Culture of Human CD34+ Cells for Lentiviral Gene Transfer -- T Cell Culture for Gammaretroviral Transfer -- Retroviral Transduction of Murine Primary T Lymphocytes -- Lentiviral Vector Gene Transfer into Human T Cells -- DNA Transposons for Modification of Human Primary T Lymphocytes -- Retroviral Gene Transfer into Primary Human Natural Killer Cells -- Lentiviral Vector-Mediated Genetic Programming of Mouse and Human Dendritic Cells -- In Situ (In Vivo) Gene Transfer into Murine Bone Marrow Stem Cells -- In Vivo and Ex Vivo Gene Transfer in Thymocytes and Thymocyte Precursors -- Design and Production of Retro- and Lentiviral Vectors for Gene Expression in Hematopoietic Cells -- Knock-Down of Gene Expression in Hematopoietic Cells -- The Use of Retroviral Vectors for tet-Regulated Gene Expression in Cell Populations -- Detection of Replication Competent Retrovirus and Lentivirus -- Release Testing of Retroviral Vectors and Gene-Modified Cells -- Copy Number Determination of Genetically-Modified Hematopoietic Stem Cells -- Tissue Procurement for Molecular Studies Using Laser-Assisted Microdissection -- Leukemia Diagnosis in Murine Bone Marrow Transplantation Models -- Humanized Mouse Models to Study the Human Haematopoietic Stem Cell Compartment -- Canine Models of Gene-Modified Hematopoiesis -- Detection of Retroviral Integration Sites by Linear Amplification-Mediated PCR and Tracking of Individual Integration Clones in Different Samples -- Retroviral Insertion Site Analysis in Dominant Haematopoietic Clones -- Tracking Gene-Modified T Cells In Vivo -- DNA Microarray Studies of Hematopoietic Subpopulations -- Quantification of Genomic Mutations in Murine Hematopoietic Cells -- Proteomics Studies After Hematopoietic Stem Cell Transplantation -- Spectral Karyotyping and Fluorescence In Situ Hybridization of Murine Cells -- Database Setup for Preclinical Studies of Gene-Modified Hematopoiesis -- The US and EU Regulatory Perspectives on the Clinical Use of Hematopoietic Stem/Progenitor Cells Genetically Modified Ex Vivo by Retroviral Vectors.

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.