Hematopoietic Stem Cell Protocols

333 Pages·2002·2.006 MB·English

by  Elaine Dzierzak, Marella de Bruijn (auth.), Christopher A. Klug PhD, Craig T. Jordan PhD (eds.)

Hematopoietic Stem Cell Protocols
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M E T H O D S I N M O L E C U L A R M E D I C I N ETM
Hematopoietic Stem
Cell Protocols
Edited by
Christopher A. Klug,
PhD
University of Alabama-Birmingham,
Birmingham, AL
and
Craig T. Jordan,
PhD
University of Kentucky,
Lexington, KY
Humana Press Totowa, New Jersey
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Main entry under title: Methods in molecular medicine™.
Hematopoietic stem cell protocols / edited by Christopher A. Klug and Craig T. Jordan.
p. ; cm. — (Methods in molecular medicine; 63)
Includes bibliographical references and index.
ISBN 0-89603-812-2 (alk. paper)
1. Hematopoietic stem cells—Laboratory manuals. I. Klug, Christopher A. II. Jordan,
Craig T. III. Series.
[DNLM: 1. Hematopoietic Stem Cells—cytology. 2. Cytological Techniques—methods.
3. Hematopoietic Stem Cells—physiology. WH 380 H48693 2001]
QP92.H4534 2001
611'.41—dc21 2001016559
Preface
The ability to highly purify and characterize hematopoietic stem cells (HSC)
from mice and humans has opened up an exceedingly rich field of basic science
research with enormous clinical potential. Many of the techniques used in stud-
ies of HSC biology have become more standardized over the last several years,
which makes it possible to compile a set of methods that can be used by both
seasoned investigators and novices in the stem cell field. We have attempted
to be as comprehensive as possible and yet focus on what we perceive to be the
most widely used approaches for studies of murine and human HSC.
This first edition of Hematopoietic Stem Cell Protocols will therefore have
some obvious omissions that were dictated by contemporary circumstances. It
is our hope that readers will feel free to contribute their personal suggestions
for further chapters as well as on how existing chapters can be improved for
future editions. We certainly expect that old approaches will be refined, new
assays will be developed, and other animal model and vector systems will be
described that will become the new gold standards for future work. Our sin-
cere thanks goes out to all of the contributors and to those in the stem cell field
that have enlarged our thinking and provided new tools to further understand
this fascinating cell type.
Christopher A. Klug, PhD
Craig T. Jordan, PhD
v
Contents
Preface .............................................................................................................v
Contributors.....................................................................................................ix
1 Isolation and Analysis of Hematopoietic Stem Cells from Mouse
Embryos
Elaine Dzierzak and Marella de Bruijn................................................1
2 The Purification of Mouse Hematopoietic Stem Cells at Sequential
Stages of Maturation
Sean J. Morrison..................................................................................15
3 Flow Cytometry and Immunoselection of Human Stem Cells
Terry E. Thomas, Sara J. Abraham, and Peter M. Lansdorp.........29
4 Method for Purification of Human Hematopoietic Stem Cells
by Flow Cytometry
Michael J. Reitsma, Brenda R. Lee, and Nobuko Uchida...............59
5 Noninvasive Measurement of Hematopoietic Stem Cell Cycling
and Turnover by In Vivo Bromodeoxyuridine Incorporation
Gillian B. Bradford and Ivan Bertoncello.........................................79
6 Isolation and Characterization of Primitive Hematopoietic Cells
Based on Their Position in the Cell Cycle
Edward F. Srour and Craig T. Jordan...............................................93
7 Hematopoietic Colony-Forming Cells
Makio Ogawa and Anne G. Livingston............................................113
8 Long-Term Culture-Initiating Cell Assays for Human
and Murine Cells
Cindy L. Miller and Connie J. Eaves...............................................123
9 The Cobblestone-Area-Forming Cell Assay
Gerald de Haan and Rob Ploemacher.............................................143
10 CFU-S: An Assay for Pluripotent Myelopoietic Stem Cells
Ernest A. McCulloch..........................................................................153
11 Intrathymic Injection for Analysis of T-Cell Progenitor Activity
Libuse Jerabek and Irving L. Weissman........................................161
vii
viii Contents
12 Quantitation of Murine and Human Hematopoietic Stem Cells by Limiting
Dilution Analysis in Competitively Repopulated Hosts
Stephen J. Szilvassy, Franck E. Nicolini, Connie J. Eaves,
and Cindy L. Miller........................................................................167
13 Ex Vivo Expansion of Human and Murine Hematopoietic
Stem Cells
Cindy L. Miller, Julie Audet, and Connie J. Eaves........................189
14 Hematopoietic Development of ES Cells in Culture
Gordon M. Keller, Saiphone Webb, and Marion Kennedy............209
15 Genetic Modification of Murine Hematopoietic Stem Cells
by Retroviruses
Christian P. Kalberer, Jennifer Antonchuk,
and R. Keith Humphries...............................................................231
16 Retroviral Transduction of FACS-Purified Hematopoietic
Stem Cells
Claudiu V. Cotta, C. Scott Swindle, Irving L. Weissman,
and Christopher A. Klug...............................................................243
17 Retroviral-Mediated Transduction and Clonal Integration Analysis
of Human Hematopoietic Stem and Progenitor Cells
Mo A. Dao and Jan A. Nolta..............................................................253
18 Production of Lentiviral Vector Supernatants and Transduction
of Cellular Targets
Richard E. Sutton...............................................................................275
19 Reverse Transcriptase-PCR Analysis of Gene Expression
in Hematopoietic Stem Cells
Donald Orlic........................................................................................287
20 Identification of Differentially Expressed Genes in Sorted Cell
Populations by Two-Dimensional Gene-Expression Fingerprinting
Alexander V. Belyavsky, Sergey V. Shmelkov,
and Jan W. M. Visser.....................................................................301
Index...........................................................................................................327
Contributors
SARA J. ABRAHAM•British Columbia Cancer Research Centre, Vancouver,
British Columbia, Canada
JENNIFER ANTONCHUK•Terry Fox Laboratory, British Columbia Cancer
Agency, Vancouver, British Columbia, Canada
JULIE AUDET • Terry Fox Laboratory, Departments of Chemical
and Bio-Resource Engineering, University of British Columbia,
Vancouver, British Columbia, Canada
ALEXANDER V. BELYAVSKY • New York Blood Center, New York, NY,
Engelhardt Institute of Molecular Biology, Moscow, Russia
IVAN BERTONCELLO • Peter MacCallum Cancer Institute, Melbourne,
Victoria, Australia
GILLIAN B. BRADFORD • Howard Hughes Medical Institute, Herman B. Wells
Center for Pediatric Research, Indiana University School of Medicine,
Indianapolis, IN
CLAUDIU V. COTTA • Department of Pathology, University of Alabama
at Birmingham, Birmingham, AL
MO A. DAO• Division of Research Immunology/Bone Marrow
Transplantation, Children’s Hospital; Departments of Pediatrics
and Craniofacial Developmental Biology, Los Angeles, CA
MARELLA DE BRUIJN•Department of Cell Biology and Genetics, Erasmus
University, Rotterdam, the Netherlands
GERALDDE HAAN•Department of Stem Cell Biology, University of Gröningen,
Gröningen, the Netherlands
ELAINE DZIERZAK • Department of Cell Biology and Genetics, Erasmus
University, Rotterdam, the Netherlands
CONNIE J. EAVES•Terry Fox Laboratory, British Columbia Cancer Agency,
StemCell Technologies Inc., Vancouver, British Columbia, Canada
R. KEITH HUMPHRIES•Terry Fox Laboratory, British Columbia Cancer
Agency, Vancouver, British Columbia, Canada
LIBUSE JERABEK• Stanford University School of Medicine, Stanford, CA
CRAIG T. JORDAN•Markey Cancer Center, University of Kentucky, Lexington, KY
CHRISTIAN P. KALBERER•Terry Fox Laboratory, British Columbia Cancer
Agency, Vancouver, British Columbia, Canada
ix
x Contributors
GORDON M. KELLER•Department of Medicine, National Jewish Medical
and Research Center, Denver, CO
MARION KENNEDY•Department of Medicine, National Jewish Medical
and Research Center, Denver, CO
CHRISTOPHER A. KLUG•Comprehensive Cancer Center, University of
Alabama-Birmingham, Birmingham, AB
PETER M. LANSDORP•Terry Fox Laboratory, British Columbia Cancer Agency,
Vancouver, British Columbia, Canada
BRENDA R. LEE•SyStemix, Inc., Palo Alto, CA
ANNE G. LIVINGSTON • Ralph H. Johnson VA Medical Center, Medical
University of South Carolina, Charleston, SC
ERNEST A. MCCULLOCH • The Ontario Cancer Institute, Princess Margaret
Hospital, Toronto, Ontario, Canada
CINDY L. MILLER•StemCell Technologies Inc., Vancouver, British Columbia,
Canada
SEAN J. MORRISON • Departments of Internal Medicine and Cell and
Developmental Biology, Howard Hughes Medical Institute and University
of Michigan, Ann Arbor, MI
FRANCK E. NICOLINI•Terry Fox Laboratory, Vancouver, British Columbia, Canada
JAN A. NOLTA • Division of Research Immunology/Bone Marrow
Transplantation, Children's Hospital, Departments of Pediatrics and
Craniofacial Developmental Biology, Los Angeles, CA
MAKIO OGAWA•Ralph H. Johnson VA Medical Center, the Medical University
of South Carolina, Charleston, SC
DONALD ORLIC•Hematopoiesis Section, Genetics and Molecular Biology
Branch, National Human Genome Research Institute, National Institute
of Health, Bethesda, MD
ROB PLOEMACHER • Institute of Hematology, Erasmus University, Rotterdam,
the Netherlands
MICHAEL J. REITSMA • StemCells Inc., Sunnyvale, CA
SERGEY V. SHMELKOV • New York Blood Center, New York, NY, Engelhardt
Institute of Molecular Biology, Moscow, Russia
EDWARD F. SROUR•Indiana Cancer Research Institute, Indiana University
School of Medicine, Indianapolis, IN
RICHARD E. SUTTON•Center for Cell and Gene Therapy, Baylor College
of Medicine, Houston, TX
C. SCOTT SWINDLE• Department of Pathology, University of Alabama
at Birmingham, Birmingham, AL
STEPHEN J. SZILVASSY • Lucille P. Markey Cancer Center, University
of Kentucky, Lexington, KY
Contributors xi
TERRY E. THOMAS•StemCell Technologies Inc., Vancouver, British Columbia,
Canada
NOBUKO UCHIDA • StemCells Inc., Sunnyvale, CA
JAN W. M. VISSER•New York Blood Center, New York, NY
SAIPHONE WEBB • Department of Medicine, National Jewish Medical
and Research Center, Denver, CO
IRVING L. WEISSMAN • Stanford University School of Medicine, Stanford, CA