Table Of ContentMethods in
Molecular Biology 1109
Meral Beksaç Editor
Bone Marrow
and Stem Cell
Transplantation
Second Edition
M M B
ETHODS IN OLECULAR IOLOGY
Series Editor
John M. Walker
School of Life Sciences
University of Hertfordshire
Hat fi eld, Hertfordshire, AL10 9AB, UK
For further volumes:
http://www.springer.com/series/7651
Bone Marrow and Stem Cell
Transplantation
Second Edition
Edited by
Meral Beksaç
Department of Hematology, School of Medicine, Ankara University, Ankara, Turkey
Editor
Meral Beksaç
Department of Hematology
School of Medicine
Ankara University
Ankara, T urkey
ISSN 1064-3745 ISSN 1940-6029 (electronic)
ISBN 978-1-4614-9436-2 ISBN 978-1-4614-9437-9 (eBook)
DOI 10.1007/978-1-4614-9437-9
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Prefa ce
Hematopoietic stem cells have been introduced to treatment of more than 40 diseases and
are being used successfully, thanks to the developments in molecular techniques and bio-
technology. Recognition of the histocompatibility complex and development of immuno-
suppressive drugs constitute the major steps leading to almost 50,000 transplants worldwide
every year. However perfect donor–recipient matching requires typing at non-HLA loci and
natural killer cell receptors too. The purpose of this book when published in 2007 was to
gather molecular methods related to stem cell transplantation from hematopoietic stem cell
molecular profi ling to in vivo tracking, donor–recipient matching and post-transplant moni-
torization as well from molecular genetics to proteomics under one roof. In the following
years, stem cells have become the core of regenerative medicine, and the scope of stem cell
transplantation has expanded from transplantation of hematopoietic stem cells to transplan-
tation of progenitors of many non-hematopoietic tissues, i.e., neurons, myocardium, bone,
and cartilage. Furthermore today, differentiated cells can be de-differentiated to behave as
embryonic stem cells. In this era, there is a need to update the fi rst edition. Thus we have
revised the continuously evolving transplantation immunology methods on HLA, minor-
HLA, and Killer Immunoglobulin-Like Receptor Typing. In addition, new chapters on
immunophenotyping and functional characterization of stem cells are included. Suffi ce to
say, the authors are eminent experts in this fi eld and we are grateful for their most valuable
contributions. Also I need to acknowledge the editorial assistance of Dr. Pinar Yurdakul.
This book may serve as a guide in the application of molecular methods for routine or
investigational purposes. As spoken by M. K. Atatürk 80 years ago in Ankara “The truest
guide in life is Science.” We hope that the book will be a reference for scientists who are
planning to start or are already advanced in stem cell transplantation studies. Every effort
towards optimal use of stem cells will bring hope to patients who are in desperate need.
Ankara, T urkey M eral Beksaç
v
Contents
Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix
1 Reporter Gene Technologies for Imaging Cell Fates in Hematopoiesis . . . . . . 1
Sophie Kusy and Christopher H. Contag
2 Flow Cytometry for Hematopoietic Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Daniela S. Krause, Michelle E. DeLelys, and Frederic I. Preffer
3 Isolation and Characterization of Mesenchymal Stem Cells . . . . . . . . . . . . . . . 47
Sedat Odabas, A. Eser Elçin, and Y. Murat Elçin
4 Methods for Functional Analysis of Stem Cells . . . . . . . . . . . . . . . . . . . . . . . . 65
Michelle Escobedo-Cousin, J. Alejandro Madrigal,
and Aurore Saudemont
5 An Overview of HLA Typing for Hematopoietic
Stem Cell Transplantation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
Katy Latham, Ann-Margaret Little, and J. Alejandro Madrigal
6 HLA Typing with Sequence-Specific Oligonucleotide
Primed PCR (PCR-SSO) and Use of the Luminex™ Technology. . . . . . . . . . . 87
Klara Dalva and Meral Beksaç
7 Sequence-Based Typing of HLA: An Improved
Group-S pecific Full-Length Gene Sequencing Approach. . . . . . . . . . . . . . . . . 101
Christina E.M. Voorter, Fausto Palusci, and Marcel G.J. Tilanus
8 Molecular Typing Methods for Minor Histocompatibility Antigens. . . . . . . . . 115
Eric Spierings
9 Natural Killer Cells and Killer-Cell Immunoglobulin-Like
Receptor Polymorphisms: Their Role in Hematopoietic
Stem Cell Transplantation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139
Jennifer Schellekens, Katia Gagne, and Steven G.E. Marsh
10 Methods of Detection of Immune Reconstitution
and T Regulatory Cells by Flow Cytometry. . . . . . . . . . . . . . . . . . . . . . . . . . . 159
Richard Charles Duggleby and J. Alejandro Madrigal
11 Molecular Methods Used for Detection of Minimal Residual
Disease Following Hematopoietic Stem Cell Transplantation
in Myeloid Disorders. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
Ahmet H. Elmaagacli
12 Molecular Methods for Detection of Minimal Residual Disease
Following Transplantation in Lymphoid and Plasma Cell Disorders. . . . . . . . . 209
Paolo Corradini and Cristiana Carniti
vii
viii Contents
13 Molecular Methods for Detection of Invasive Fungal Infections
and Mycobacteria and Their Clinical Significance in Hematopoietic
Stem Cell Transplantation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 239
Pinar Yurdakul and Sule Colakoglu
14 Post-transplant Monitoring of Chimerism by Lineage-S pecific Analysis . . . . . . 271
Sandra Preuner and Thomas Lion
15 Urinary Proteomics Employing Capillary Electrophoresis Coupled
to Mass Spectrometry in the Monitoring of Patients After
Stem Cell Transplantation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 293
Eva M. Weissinger, William Mullen, and Amaya Albalat
Erratum. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . E1
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 307
Contributors
AMAYA ALBALAT • BHF Glasgow Cardiovascular Research Centre , U niversity of Glasgow ,
Glasgow, U K
J. ALEJANDRO MADRIGAL • Anthony Nolan Research Institute, University College London ,
London, U K
MERAL BEKSAÇ • Department of Hematology , School of Medicine, Ankara University ,
Ankara, Turkey
CRISTIANA C ARNITI • Division of Hematology, Bone Marrow Transplantation Unit,
Fondazione IRCCS Istituto Nazionale Tumori Milano, University of Milano ,
Milan, I taly
SULE C OLAKOGLU • Department of Microbiology and Clinical Microbiology , B askent
University Faculty of Medicine Adana Hospital , A dana, Turkey
CHRISTOPHER H. CONTAG • Department of Pediatrics, S tanford University School of
Medicine , S tanford , C A , USA
PAOLO CORRADINI • Division of Hematology, Fondazione IRCCS Istituto Nazionale
Tumori Milano, University of Milano , Milan, I taly
KLARA DALVA • Department of Hematology, Tissue Typing Laboratories, I bni Sina Hospital,
Ankara University , A nkara , T urkey
MICHELLE E. D ELELYS • Department of Pathology, Massachusetts General Hospital , B oston ,
MA , U SA
RICHARD CHARLES D UGGLEBY • Anthony Nolan Research Institute, Royal Free Hospital ,
London, U K
Y. M URAT ELÇIN • Faculty of Sciences and Stem Cell Institute, Ankara University , Ankara,
Turkey
A. E SER ELÇIN • Faculty of Sciences and Stem Cell Institute, Ankara University , Ankara,
Turkey
AHMET H. E LMAAGACLI • Department of Bone Marrow Transplantation , U niversity
Hospital of Essen , H ufelandstr, G ermany
MICHELLE ESCOBEDO-COUSIN • Anthony Nolan Research Institute, University College
London , L ondon, UK
KATIA G AGNE • Etablissement Français du Sang and Université de Nantes,
Immunovirologie et Polymorphisme Génétique , Nantes, F rance
DANIELA S. KRAUSE • Department of Pathology, M assachusetts General Hospital ,
Boston, MA , U SA
SOPHIE K USY • Department of Pediatrics, Stanford University School of Medicine ,
Stanford , C A , USA
KATY L ATHAM • Anthony Nolan Research Institute, University College London , L ondon, UK
THOMAS LION • Children’s Cancer Research Institute (CCRI) , Vienna, A ustria
ANN-MARGARET L ITTLE • Histocompatibility and Immunogenetics, Greater Glasgow
and Clyde, Gartnavel General Hospital , Glasgow, UK
STEVEN G . E. MARSH • Laboratoire HLA-EFS , Nantes, F rance
ix
