Cell Separation Methods And Applications

Page i
Cell Separation Methods and Applications
edited by
Diether Recktenwald
AmCell Corporation
Sunnyvale, California
Andreas Radbruch
Deutsches Rheuma-Forschungszentrum Berlin
Berlin, Germany

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Library of Congress Cataloging-in-Publication Data
Cell separation methods and applications / edited by Diether Recktenwald, Andreas
Radbruch.
p. cm.
Includes bibliographical references and index.
ISBN 0-8247-9864-3
1. Cell separation. I. Recktenwald, Diether. II. Radbruch, A. (Andreas)
QH585.5.C44C435 1997
571.6—dc21 97 -33113
CIP
The publisher offers discounts on this book when ordered in bulk quantities. For more information, write to
Special Sales/Professional Marketing at the address below.
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Copyright © 1998 by MARCEL DEKKER, INC. All Rights Reserved.
Neither this book nor any part may be reproduced or transmitted in any form or by any means, electronic or
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system, without permission in writing from the publisher.
MARCEL DEKKER, INC.
270 Madison Avenue, New York, New York 10016
http://www.dekker.com
Current printing (last digit):
10 9 8 7 6 5 4 3 2 1
PRINTED IN THE UNITED STATES OF AMERICA

Page iii
FOREWORD
It should not come as a surprise that the fields of developmental biology as an intellectual discipline and cell
separation as a technological discipline have grown in an interdependent fashion. Just as the cell is the unit of
biological organization, cells are organized into groupings from the most mature effector cells back to the most
primitive progenitors-the stem cells. Stem cells and progenitor cells have the capacity of being clonogenic
precursors for large numbers of progeny; stem cells are the most primitive subset and are defined as the cells
that can both self-renew and at the single-cell level give rise to progeny of several different mature cells.
Progenitors may be multipotent or oligopotent and can be distinguished from stem cells by their lack of selfrenewal
capacity. The generation and regeneration of all tissues in the organs in the body depend on the actions
of stem and progenitor cells. When we think of generation, we think of those events that occur in embryonic
and fetal life to give rise to the organ systems; and when we think of regeneration, we think largely of repair
systems, which, in the end, can or should be exploited clinically to replace, wholly or in part, tissue and organ
systems that are damaged or are defective.
Identification of the stem and progenitors cells involved in generation or regeneration of a particular organ
system or tissue concerns fairly rare populations. When one is dealing with regeneration of tissue or organ
systems in a clinical circumstance where diseased or malignant cells may contaminate that organ system,
practical regeneration can occur only if the stem/progenitor cells

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are purified nearly to homogeneity and do not contain contaminating diseased cells. To do this not only in
animal models but in humans, one must be thinking of separation technologies that not only are high fidelity in
terms of identifying and isolating rare populations, but also are on a scale that is large enough to deliver
adequate numbers of stem/progenitor cells for clinically effective and rapid regeneration.
As an immunologist, I must also add that large-scale identification and separation of subsets of cells of the
immune system for experiment and clinical treatment involve clonogenic cells of another sort-lymphocytes that
respond to antigen by clonal expansion and differentiation into memory or effector cells. Because most
methods that allow the culture and expansion of these important potential effector cells also alter their life-span
and homing properties to the extent that normal regeneration of the immune system cannot occur, again largescale
high-fidelity separations are required.
Within this exciting volume two masters of cell separation technologies, Diether Recktenwald and Andreas
Radbruch, have gathered together the most accomplished researchers at the leading edge of several cell
separation technologies. It is fitting that there is input from the commercial arena, as the development of these
technologies as research tools and as clinical scale separation devices can only come through entrepreneurial
and commercial efforts. These articles collectively provide the technological and scientific basis for advancing
cell separation and identification technologies, and I commend them to you as the state of the art just prior to a
new era when this marriage of cell, developmental, and cell separation technologies is about to transform
medicine and, at the same time, reveal new insights into the cell and molecular biology that allows stem and
progenitor cells to be just that.
IRVING WEISSMAN, PH.D.
DEPARTMENT OF PATHOLOGY
AND DEVELOPMENTAL BIOLOGY
STANFORD UNIVERSITY
STANFORD, CALIFORNIA

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PREFACE
The need for ever more powerful methods in cell separation has grown tremendously with the identification of
many specialized cell subsets because of rapid progress in cell biology, immunology, and molecular biology.
This book on Cell Separation Methods and Applications describes all of the important methods for the
analytical and preparative isolation of specific populations of biological cells. In Chapter 1, Esser includes
basic methods such as lytic removal of cell subsets. These methods were developed several decades ago and are
still used, primarily as a prepurification step for the preparation of certain common mammalian hematopoietic
cell populations for studies in biochemistry, cell biology, immunology, molecular biology, and clinical
research.
The newest developments in methods using endogenous physical cell properties, such as cell size and cell
density, are described in two chapters by leaders in the field of centrifugal elutriation (Van Vlasselaer and coworkers)
and density gradient separations (Figdor and colleagues).
All of the more specific cell separation methods, based on monoclonal antibodies against cell surface markers,
are described by academic and industrial pioneers working on the perfection of the techniques. The methods
include polystyrene immunoaffinity devices (Lebkowski and co-workers), biotin avidin
immunochromatography (Heimfeld and co-workers), high-density particles (Kenyon and co-workers),
complement-based cell lysis (Gee), various immunomagnetic methods for cell sorting (Kantor and co-workers),

Page vi
magnetophoresis (Hausmann and co-workers), and fluorescence-activated cell sorting based on flow cytometry
(Hoffman and Houck)
The book also describes the basic technical principles of the respective cell separation methods in detail.
Typical examples with performance data are discussed, and the limitations of the methods are outlined.
Applications in basic research and medicine and an outlook on future applications are reviewed. For ease of
understanding, flowcharts, figures, and tables support the text in all chapters.
Finally, the book includes several chapters on important applications of combinations of cell separation
techniques, including research in molecular and cellular biology and genetics (Siebenkotten and co-workers)
and the clinical isolation of stem and progenitor cells for cell therapy of cancer and other diseases (Hassan and
co-workers).
To aid the researcher with the design of cell separation projects, there is an appendix with cell properties for
cell separations and one with CD antigens updated with results from the February 1996 leukocyte antigen
workshop in Osaka.
This book will help those working with cellular preparations to select an optimal cell separation approach, and
it will provide a detailed understanding of the possibilities and limitations of cell purification for those involved
in clinical cellular therapy for transplantation, cancer, and AIDS, or with gene therapy for a variety of genetic
defects.
We hope that the methods explained in this book will contribute to major advances in cellular therapy and
diagnostics, which, in turn, will lead to refinements in methodology.
We would like to acknowledge those who provided valuable support in the preparation of the book, including
Larry Transue, technical typist; and Kathryn Rubenstein and her co-workers, graphics. Without MaryAnn
Foote, Ph.D., this book would not exist. Without her energy and skill in organizing the book and keeping it on
schedule, the project would not have succeeded.
DIETHER RECKTENWALD, PH.D.
ANDREAS RADBRUCH, PH.D.

Pa
CONTENTS
Foreword
(Irving Weissman)
Preface
Contributors
Part 1: Background
1. Historical and Useful Methods of Preselection and Preparative Scale Sorting
Charlotte Esser
Part II: Physical Methods of Separation
2. New Approaches in Density Gradient Separation Using Colloidal Silica Gradients in the
Processing of Human Hematopoietic Progenitor Cells
Peter Van Vlasselaer, Varghese C. Palathumpat, George Strang, and Michael H. Shapero
3. Centrifugal Elutriation: A Powerful Separation Technique in Cell Biology, Immunology, and
Hematology
Carl G. Figdor, Frank Preijers, Richard Huijbens, Paul Ruijs, Theo J. M. de Witte, and Willy S.
Bont

Pag
Part III: Antibody-Based Methods
4. Isolation, Activation, Expansion, and Gene Transduction of Cell-Based Therapeutics Using
Polystyrene Immunoaffinity Devices
Jane S. Lebkowski, Dewey J. Moody, Ramila Philip, Lisa Schain, Sohel Talib, Rukmini Pennathur-
Das, David A. Okrongly, and Thomas B. Okarma
5. The CEPRATE® SC System: Technology, Clinical Development, and Future Directions
Shelly Heimfeld, Karen Auditore-Hargreaves, Mark Benyuenes, Michael Emde, Cindy Jacobs,
Mark Jones, Nicole Provost, Grant Risdon, and Joe Tarnowski
6. High-Density Particles: A Novel, Highly Efficient Cell Separation Technology
Norma S. Kenyon, Robert K. Zwerner John G. Gribben, Lee M. Nadler, Camillo Ricordi, and
Thomas R. Russell
7. Antibody- and Complement-Mediated Cell Separation
Adrian P. Gee
Part IV: Magnetic Methods
8. Magnetic Cell Sorting with Colloidal Superparamagnetic Particles
Aaron B. Kantor, Ian Gibbons, Stefan Miltenyi, and Jürgen Schmitz
9. Immunomagnetic Cell Separation Using Antibodies and Superparamagnetic Microspheres
Adrian P. Gee
10. Free-Flow Magnetophoresis: Continuous Immunomagnetic Sorting of Cells and Organelles by
Magnetic Deviation and Focusing
Michael Hausmann, Roland Hartig, Hans-Georg Liebich, Georg H. Lüers, Armin Saalmüller,
Reinhard Teichmann, and Christoph Cremer

Pa
Part V: Flow Cytometry
11. Cell Separation Using Flow Cytometric Cell Sorting
Robert A. Hoffman and David W. Houck
Part VI: Special Applications
12. Employing Surface Markers for the Selection of Transfected Cells
Gregor Siebenkotten, Katja Petry, Ute Behrens-Jung, Stefan Miltenyi, and Andreas Radbruch
13. CD34 + Cell Sorting and Enrichment: Applications in Blood Banking and Transplantation
Hassan T Hassan, K. Gutensohn, A. R. Zander, and P. Kuehnl
Appendixes
A. Cellular Properties for Cell Separation
B. CD Antigen Designations
Glossary
Index

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INDEX
A
N-acetyl galactosamine, 63
Acquired immune deficiency syndrome (AIDS), 62, 81
ADA, see Adenosine deaminase
Adeno-associated virus (AAV), 77 (figure)
plasmids, 75
Adenosine deaminase (ADA), 81, 98
Adhesion molecules, 73 (table), 285
AIDS, see Acquired immune deficiency syndrome
Albumin, 190
Analysis, residual cell, 188, 198
Anemia,
aplastic, 290
refractory, 290
Antibody,
fluorescein-conjugated, 67, 126
fluorochrome-conjugated, 66, 79 (figure)
OKT3, 138
phycoerythrin-conjugated, 66, 122 (figure), 126
use in cell sorting, 1, 5, 61, 62, 63, 67, 80, 88, 90, 96, 103, 104, 106, 115, 133–146, 154, 160, 161, 175–
176, 177, 179, 184, 186, 188, 198, 210, 225, 230, 238, 243, 258, 261, 278
Antigen, use in cell sorting, 1, 65, 68, 82, 88, 155, 186, 188, 243, 273, 274
Apoptosis, 249, 259–260

Avidin, columns, 11 (table), 88–89, 89 (figure)
B
B cells, 2, 7, 8, 10, 28 (figure), 34, 51 (figure), 163 (figure), 164, 272, 273, 274
depletion of, 34
Bags, gas-permeable, 69
Beads,
anti-immunoglobulin, 183, 188, 191
avidin, 91, 92 (figure), 156, 161, 179, 210
coating protocols, 189
Dynal, 176, 177 (figure), 181, 210, 232
fluorochrome, 156
hapten, 156, 164
incubating protocols, 191
latex, 220, 221, 221 (table)
magnetic, 154, 155, 220, 221, 229
polyacrylamide, 88, 89, 90, 91
streptavidin, 188, 228
Biohazard, 248, 265
Biotin, 228
columns, 88–89, 89 (figure), 156, 162
Blood,
cord, 88, 98, 117, 284, 284 (table)
murine, 2
Bone marrow cells, 19, 52, 69, 70 (figure), 75, 90, 246, 258, 261, 284
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mononuclear cells of, 63, 65, 66 (table), 67, 68 (figure), 74 (figure)
transplantation of, 52–53, 62, 80–81, 94, 95, 104, 108, 111, 114, 121, 146, 166, 261, 285
vertebral, use of, 114–117, 118 (figure), 119 (figure)
Borohydride reduction, 156
C
Calcium, 239
Cancer, 82, 289
breast, 94, 95, 96, 98, 170, 182, 186, 289
lung, 289
neuroblastoma, 182
ovarian, 72, 73
small-cell lung, 182
Catcher tube sorting, 248–249, 248 (figure)
Cell cycle, separation of cells according to, 51, 258
Cell selection,
positive, 133, 158, 159–160, 162–166, 181, 182 (figure), 200, 201
negative, 133, 158, 159–160, 162, 181, 182 (figure), 200
two-step, 200–201
Cell selectors,
Ceprate SC, 87–101
MacroCELLector, 62–63, 78
RPR CELLector, 62–66, 68–70, 72–76
Cell separation,
direct, 191

direct versus indirect, 183–186, 184 (figure), 185 (figure)
indirect, 187 (figure), 191
using flow cytometry for, 237–265
Cell sorting,
efficacy of, 252–254
protocols, 2–10, 11 (table), 20, 63
purity of, 252–254
Cell types,
also see specific names
CD2, 96
CD3, 51 (figure), 64, 161, 165 (figure), 169, 187
CD4, 64, 66, 81, 99, 112 (figure), 126, 162, 273 (figure), 275, 276, 278 (figure)
CD5, 64, 66, 71, 80, 161, 164, 170 (figure)
CD8, 64, 66, 67, 72, 74 (figure), 76 (figure), 78, 80 (figure), 81, 126, 272
CD15, 115, 117
CD19, 99, 164, 165 (figure), 222, 224, 284
CD20, 27
CD25, 272
CD34, 16, 19, 20–24, 25 (figure), 28 (figure), 29, 30 (figure), 34, 48, 50, 52, 53, 63, 64, 66, 67, 68 (figure),
70, 75, 77 (figure), 91, 94, 98–99, 103, 115, 117, 169, 170 (figure), 171 (table), 183, 190, 258, 279, 283–
290
CD38, 64, 67, 68 (figure)
CD45, 25 (figure), 30 (figure), 162
CD56, 99
gene-modified, 77–78, 261
H-2K k , 273, 274, 274 (figure), 276, 277 (figure)
MOLT-4, 107

Cell volume, electronic measurement of, 245
Centrifugal elutriation, 43–54
basic principles of, 44–46
chambers, types used in, 47
closed systems, 48
computer-assisted (CACE), 50
reducing sample size by, 48
use of flow systems in, 47
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Centrifugation, 24, 25, 27, 28 (figure), 52
CFU, see Colony-forming units
Chemotherapy, 290
Chimerism, 113
Chromatography, 159
Chromium-release assay, 26, 74 (figure), 142
Chromosomes, 226–229, 228 (figure), 262
Chymopapain, 195
Clinical use, 48, 52, 53, 78, 80–82, 87, 104, 210, 233, 263, 283–290
Cobalt, 155
COBE 2991, 27
Colony-forming units/cells (CFU/C), 19, 24, 34, 283
Complement, 9, 10 (table), 11 (table), 176, 201
Cotton wool, 5, 6 (figure and table), 11 (table)
Cyanogen bromide, 157
Cytokeratin, 166, 167 (figure)
Cytokines, see specific names
Cryopreservation, 289, 290
Cytostatic drugs, 52
D
Dendritic cells, 51, 53, 82, 103, 104, 126, 162, 163 (figure)
Density-adjusted cell sorting (DACS), 29–34
Density gradient material,
characteristics of, 17

DETACHaBEAD, 195
Diaminohexane, 157
Dimethylsulfoxide (DSMO), 29, 93 (figure), 114
Diphtheria toxin, 134
Dispase, 195
DNA,
analysis, 126, 228, 239, 249
transfection, 271–279
DSMO, see Dimethylsulfoxide
Dyspnea, 29
E
Electromagnets, 216–217
Electronic gate dissemination, 2
Elutriation, 106
Endocrine cells, 225
Eosin, 145
Eosinophils, 126
Epithelial cells, 51, 225, 226 (figure)
Erythrocytes, 2, 3, 50, 115, 117, 162, 164 (figure), 223, 224 (table), 264
lysis of, 2
Erythroid burst-forming cells (E-BFC), 70
Erythropoietin (EPO), 290
Ex vivo expansion, 98, 289, 290
F
FACS, see Fluorescence-activated cell sorting

Fc receptor, 157, 160, 181
Ficoll, 1, 2, 3, 4, 4 (figure and table), 11 (table), 15, 24, 25, 107, 109, 111, 117, 142, 259
Flow cytometry, 67, 145, 153, 222, 225, 237–265, 241 (figure), 272, 274 (figure)
one-parameter, 50, 286
multiparameter, 239–245, 244 (figure)
two-parameter, 50, 67, 116 (figure)
Fluidic-switching sorting, 249–250, 249 (figure)
Fluorescence cell sorting (FACS), 1, 2, 15, 33 (figure), 158, 159, 163 (figure), 165 (figure), 167 (figure), 170
(figure)
Fluorochromes, 237, 238–239
Forward scatter, 240, 242, 245 (figure)
G
Gastrin cells, 224–225
Gaucher's disease, 98
Gene,
CAT, 75, 273
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env, 72
gag/pol, 72
mdr, 273
nef, 72
rev, 81
therapy, 81, 98, 104, 166, 277, 279, 289
tk, 278
transduction, 63
transfer, 75
Glutaraldehyde, 225
Glycophorin, 109, 117, 120 (figure), 162
Graft-versus-host disease (GVHD), 26, 27, 28, 52, 81, 96, 99, 103, 288–289
Graft-versus-leukemia (GVL), 26, 288–289
Granulocytes, 3, 109, 223
Granulocyte-macrophage colony-forming cell (GM-CFC), 70, 95, 95 (table), 96 (table)
GVHD, see Graft-versus-host disease
GVL, see Graft-versus-leukemia
H
Hematopoietic progenitor cells, 18, 19, 20–23, 25, 27, 29, 50
Hemoglobinuria, 29
Heparin, 143
Hepatocytes, 51
High-density particles, separation using, 103–129
HIV, see Human immunodeficiency virus

Human immunodeficiency virus, 98
gene, 72, 74, 75
infection, 248
N-hydroxy-succinimide, 157
I
IFN, see interferon
IL, see interleukin
Immunoabsorption, 258
Immunoaffinity, 134
Immunoglobulin, presence of, 67
Immunomagnetic techniques, 134, 135, 139, 153–171, 175–201, 209–233, 258, 273, 275
continuous, 211–233
Immunotherapy, 53, 81, 88, 101
Immunotoxin, 176, 201
Integrins, 285
Interferon (IFN),
α, 61
γ, 53, 61
Interleukin (IL),
-2, 61, 73, 76, 81, 272
-3, 75, 99, 285
-6, 75, 99
Internet, 238
Investigational device exemption (IDE), 140
Investigational new drug (IND), 140
Iron, 155

dextran, 155–156, 156 (table), 159
oxide, 210
Isothiocyanate, 157, 165 (figure), 170 (figure), 273, 277 (figure)
K
Keratinocytes, 51
Kupffer cells, 51
L
Laser, 239–240, 240 (figure), 242, 242 (figure), 247
tweezers, 246
Lectin, 62, 179
L-leucine methyl ester, 1
Leukemia, 285–286
acute lymphocytic (ALL), 81, 286
acute myelogenous (AML), 81, 286
chronic myelogenous (CML), 81, 286
Leukocytes, 2, 53, 162, 190, 285
Lipofection, 81
Lipopolysaccharide, 270
Liposomes, 75
Liver transplantation, 113
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Long-term culture-initiating cell (LTCIC), 283
Lymphocytes, 1, 18, 47, 50, 51 (figure), 100 (figure), 103, 109, 126, 139, 222, 224, 225, 243, 245, 258, 279
Lymphoma, 124–125, 135, 182
M
Macrophages, 2, 8, 190
MACS, see Magnetic cell sorting
Magnetic cell separation, 192–195
Magnetic cell sorting (MACS), 1, 2, 153–171, 272, 274, 274 (figure), 275, 276
Magnetic filter, 211, 224
Major histocompatibility class (MHC)
I, 273 (figure)
II, 10, 276, 278 (figure)
Markers,
activation, 73 (table)
other, 73 (table)
surface, 271–279
Medium, Dulbecco's Modified Eagle (DMEM), 63, 75
Melanoma-associated antigens, 53
Methacrylate, 155
Methylcellulose, 15
Metrizamide, 15
MHC, see Major histocompatiblity class
Monocytes, 1, 18, 43, 50, 51 (figure), 53, 162, 163 (figures), 223
mpl ligand, 285, 290

Myelocytes, 70
N
Natural killer (NK) cells, 1, 18, 26, 27 (figure), 29, 162, 163 (figure)
Neodymium-iron-boran magnets, 180
Nerve growth factor receptor, 78
Neutrophils, 70
recovery of, after transplantation, 34, 94, 290
NHL, see Non-Hodgkin's lymphoma
Nickel, 104, 105 (figure), 016, 107, 107 (table), 108
Nitrocellulose filters, 259
Non-Hodgkin's lymphoma, 19
NK cells, see natural killer cells
Nuclei, separation of, 250
Nylon wool, 2, 8, 11 (table), 190
O
Optical scanning, 212, 220, 229 (figure), 237, 242 (figure)
O-sialoglycoprotease, 195
Osteoblasts, 18
P
Pancreatic islets, 249, 262–263
Panning, 1, 5, 7 (figure), 8 (table), 11 (table), 134, 272, 275
Parasites, 249
PBMC, see Peripheral blood mononuclear cells
PBPC, see Peripheral blood progenitor cells
PEG, see Polyethylene glycol

Percoll, 3, 11 (table), 17–18, 19, 24, 45
Perfusion system, 212
Peripheral blood mononuclear cells (PBMC), 3, 5 (table), 50, 62, 64, 66 (table), 72, 76, 161 (figure), 162, 163
(figure), 165 (figure), 167 (figure)
Peripheral blood progenitor cells (PBPC), 63–64, 87, 90, 284
transplantation of, 94, 286–290
Peroxisomes, 230, 231 (figure)
Phagocytosis, 190, 191
Phycoerythrin, 222, 273 (figure)
Phytohemagglutinin, 65, 74, 76, 81
Plasma membrane, 239, 245, 250
Platelets, 28 (figure), 64, 111, 115 (figure), 223
recovery of, 34, 94
Poisson statistics, 252–253
Polyacrylamide, 88
Polyethylene glycol (PEG), 17
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Polymerase chain reaction (PCR), 106, 124, 125, 135, 145, 198, 249, 259
Polysaccharide, 155
Polystyrene, immunoaffinity devices, 61–82, 176–201, 210
Polyvinylpyrrolidone (PVP), 17
Prion, 279
Promyelocytes, 70
Pulmonary edema, 29
PVP, see Polyvinylpyrrolidone
Q
QBEND10, 283
R
Rare cells, 210, 213, 224 (table), 249, 264, 271, 275
Recombinant human granulocyte colony-stimulating factor (rHuG-CSF), 19, 20, 23, 27, 28 (figure), 30 (figure),
34, 50, 52, 94, 94 (table), 97, 97 (table), 117, 120 (figure), 284 (table), 289
Recombinant human granulocyte-macrophage colony-stimulating factor (rHuGM-CSF), 51, 52, 53, 289
Recombinant human interleukin (rHuIL)
-1, 53, 69
-2, 65, 72, 81
-3, 69, 289, 290
-7, 72
-11, 290
Recombinant human stem cell factor (rHuSCF), 69, 290
Recovery of cells, 257
Red blood cell, see Erythrocytes
Renal cell carcinoma, 81

cell line, 69
rHuG-CSF, see Recombinant human granulocyte colony-stimulating factor
rHuGM-CSF, see Recombinant human granulocyte-macrophage colonystimulating factor
rHuIL, see Recombinant human interleukin
rHuSCF, see Recombinant human stem cell factor
Ricin, 134
RNA, 239
Rosette, 1, 11 (table), 179–180, 181, 192, 193, 195, 211, 275
S
Samarium-cobalt magnets, 180
SBA, see Soybean agglutinin
Sephadex G-10, 11 (table)
Side scatter, 240, 242, 245 (figure), 286
Silica, 11 (table)
separation with, 15–36
Signal transduction, 71
SK-BR3 breast cancer cell line, 34, 35 (figure)
Sorters,
cell killing, 250
droplet, 243 (figure), 246–247
enclosed, 247–250
Southern blot analysis, 276
Soybean agglutinin, 63
Sperm, 18, 51, 264–265
Spleen cells, murine, 2, 3 (table)
Steel wool, 157–158, 162, 211

Stem cell factor (SCF), 75, 99
Stokes Law, 16
SU-DHK4 lymphoma cell line, 34, 35 (figure)
Sulfhydryl, 157
T
T47D epithelial tumor cell line, 166, 167 (figure)
T cells, 1, 7, 8, 9 (table), 27, 32 (figure), 51 (figure), 52, 53, 75, 160, 161, 162, 163 (figure), 245, 272, 288
depletion of, 34, 35, 63, 64, 65, 67,
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80–81, 88, 95, 96, 97 (figure), 98, 104, 109–110, 169, 177, 183, 190, 199, 222 (table)
expansion of, 81
receptor, 72, 73 (table)
Thalassemia, 290
Thrombocytopenia, 290
Thymocytes, 51
Trypan blue, 10 (table), 145, 225
Trypsin, 195
Tumor cells, 166, 258 (table)
depletion of, 34, 35 (figure), 53, 88, 121, 169, 177
Tumor-infiltrating lymphocytes (TIL), 64, 69 (figure), 75, 81, 103
V
Vaccinia virus constructs, 73, 74 (figure)
W
White blood cells, see specific names
X
Xanthan gum, 263
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