Preface ........................................................ v
Contributors ................................................ xvii
1 Immunomagnetic Enrichment of Human and Mouse Hematopoietic
Stem Cells for Gene Therapy Applications ..................... 1
Guillermo Guenechea, Jose C. Segovia, and Juan A. Bueren
2 Isolation of Human and Mouse Hematopoietic Stem Cells ....... 13
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 .......................................... 23
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 ........................... 33
Tulin Budak-Alpdogan and Isabelle Rivière
5 Short-Term Culture of Human CD34+Cells for Lentiviral
Gene Transfer ............................................... 59
Francesca Santoni de Sio and Luigi Naldini
6 T Cell Culture for Gammaretroviral Transfer ................. 71
Sebastian Newrzela, Gunda Brandenburg, and Dorothee
von Laer
7 Retroviral Transduction of Murine Primary T Lymphocytes ..... 83
James Lee, Michel Sadelain, and Renier Brentjens
8 Lentiviral Vector Gene Transfer into Human T Cells .......... 97
Els Verhoeyen, Caroline Costa, and Francois-Loic Cosset
9 DNA Transposons for Modification of Human Primary
T Lymphocytes .............................................. 115
Xin Huang, Andrew Wilber, R. Scott McIvor, and Xianzheng
Zhou
10 Retroviral Gene Transfer into Primary Human Natural
Killer Cells ............................................... 127
Evren Alici, Tolga Sutlu, and M. Sirac Dilber
11 Lentiviral Vector-Mediated Genetic Programming of Mouse
and Human Dendritic Cells .................................. 139
Renata Stripecke
12 In Situ (In Vivo) Gene Transfer into Murine Bone Marrow
Stem Cells ................................................. 159
Dao Pan
13 In Vivo and Ex Vivo Gene Transfer in Thymocytes and
Thymocyte Precursors ....................................... 171
Oumeya Adjali, Amelie Montel-Hagen, Louise Swainson,
Sophie Marty, Rita Vicente, Cedric Mongellaz, Chantal
Jacquet, Valérie Zimmermann, and Naomi Taylor
14 Design and Production of Retro- and Lentiviral Vectors
for Gene Expression in Hematopoietic Cells ................. 191
Axel Schambach, William P. Swaney, and Johannes C.M. van
der Loo
15 Knock-Down of Gene Expression in Hematopoietic Cells ....... 207
Michaela Scherr, Letizia Venturing and Matthias Eder
16 The Use of Retroviral Vectors for tet-Regulated Gene
Expression in Cell Populations ............................. 221
Rainer Löw
17 Detection of Replication Competent Retrovirus and
Lentivirus ................................................. 243
Lakshmi Sastry and Kenneth Cornetta
18 Release Testing of Retroviral Vectors and Gene-Modified
Cells ...................................................... 265
Diana Nordling, Anne Kaiser, and Lilith Reeves
19 Copy Number Determination of Genetically-Modified
Hematopoietic Stem Cells ................................... 281
Todd Schuesler, Lilith Reeves, Christofvon Kalle, and
Elke Grassman
20 Tissue Procurement for Molecular Studies Using Laser-
Assisted Microdissection ................................... 299
Ulrich Lehmann and Hans Kreipe
21 Leukemia Diagnosis in Murine Bone Marrow Transplantation
Models ..................................................... 311
Zhixiong Li, Ute Modlich, and Anjali Mishra
22 Humanized Mouse Models to Study the Human
Haematopoietic Stem Cell Compartment ....................... 331
Dominique Bonnet
23 Canine Models of Gene-Modified Hematopoiesis ............... 341
Brian С. Beard and Hans-Peter Kiem
24 Detection of Retroviral Integration Sites by Linear
Amplification-Mediated PCR and Tracking of Individual
Integration Clones in Different Samples .................... 363
Manfred Schmidt, Kerstin Schwarzwaelder, Cynthia С.
Bartholomae, Hanno Glimm, and Christofvon Kalle
25 Retroviral Insertion Site Analysis in Dominant
Haematopoietic Clones ...................................... 373
Olga S. Kustikova, Ute Modlich, and Boris Fehse
26 Tracking Gene-Modified T Cells In Vivo ..................... 391
Alessandra Recchia and Fulvio Mavilio
27 DNA Microarray Studies of Hematopoietic Subpopulations ..... 403
Karin Pike-Overzet, Dick de Ridder, Tom Schonewille,
and Prank J.T. Staal
28 Quantification of Genomic Mutations in Murine
Hematopoietic Cells ........................................ 423
Hartmut Geiger, David Schleimer, Kalpana J. Nattamai,
and Jan Vijg
29 Proteomics Studies After Hematopoietic Stem Cell
Transplantation ............................................ 437
Eva M. Weissinger, Petra Zürbig, and Arnold Ganser
30 Spectral Karyotyping and Fluorescence
In Situ Hybridization of Murine Cells ...................... 453
Cornelia Rudolph and Brigitte Schlegelberger
31 Database Setup for Preclinical Studies of Gene-Modified
Hematopoiesis .............................................. 467
Brenden Balcik, Elke Grassman, and Lilith Reeves
32 The US and EU Regulatory Perspectives on the Clinical
Use of Hematopoietic Stem/Progenitor Cells Genetically
Modified Ex Vivo by Retroviral Vectors ..................... 477
Carolyn A. Wilson and Klaus Cichutek
Index ......................................................... 489
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