cell separation methods and applications

Page iiLibrary of Congress Cataloging-in-Publication Data Cell separation methods and applications / edited by Diether Recktenwald, Andreas This book is printed on acid-free paper.. To d

Andreas Radbruch

Deutsches Rheuma-Forschungszentrum Berlin

Berlin, Germany

Page ii

Library of Congress Cataloging-in-Publication Data

Cell separation methods and applications / edited by Diether Recktenwald, Andreas

This book is printed on acid-free paper

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 mechanical, including photocopying, microfilming, and recording, or by any information storage and retrieval system, without permission in writing from the publisher

MARCEL DEKKER, INC

270 Madison Avenue, New York, New York 10016

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 self-renewal 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

Page ivare 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 large-scale 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 PATHOLOGYAND DEVELOPMENTAL BIOLOGYSTANFORD UNIVERSITY

STANFORD, CALIFORNIA

Page v

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 co-workers) 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 vimagnetophoresis (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

1 Historical and Useful Methods of Preselection and Preparative Scale Sorting

Part II: Physical Methods of Separation

Peter Van Vlasselaer, Varghese C Palathumpat, George Strang, and Michael H Shapero

2 New Approaches in Density Gradient Separation Using Colloidal Silica Gradients in the

Processing of Human Hematopoietic Progenitor Cells

Carl G Figdor, Frank Preijers, Richard Huijbens, Paul Ruijs, Theo J M de Witte, and Willy S

Bont

3 Centrifugal Elutriation: A Powerful Separation Technique in Cell Biology, Immunology, and

Hematology

Part III: Antibody-Based Methods

Jane S Lebkowski, Dewey J Moody, Ramila Philip, Lisa Schain, Sohel Talib, Rukmini Pennathur

-Das, David A Okrongly, and Thomas B Okarma

4 Isolation, Activation, Expansion, and Gene Transduction of Cell-Based Therapeutics Using

Polystyrene Immunoaffinity Devices

Shelly Heimfeld, Karen Auditore-Hargreaves, Mark Benyuenes, Michael Emde, Cindy Jacobs,

Mark Jones, Nicole Provost, Grant Risdon, and Joe Tarnowski

5 The CEPRATE® SC System: Technology, Clinical Development, and Future Directions

Norma S Kenyon, Robert K Zwerner John G Gribben, Lee M Nadler, Camillo Ricordi, and

Thomas R Russell

6 High-Density Particles: A Novel, Highly Efficient Cell Separation Technology

Adrian P Gee

7 Antibody- and Complement-Mediated Cell Separation

Part IV: Magnetic Methods

Aaron B Kantor, Ian Gibbons, Stefan Miltenyi, and Jürgen Schmitz

8 Magnetic Cell Sorting with Colloidal Superparamagnetic Particles

Adrian P Gee

9 Immunomagnetic Cell Separation Using Antibodies and Superparamagnetic Microspheres

Michael Hausmann, Roland Hartig, Hans-Georg Liebich, Georg H Lüers, Armin Saalmüller,

Reinhard Teichmann, and Christoph Cremer

10 Free-Flow Magnetophoresis: Continuous Immunomagnetic Sorting of Cells and Organelles by

Magnetic Deviation and Focusing

Part V: Flow Cytometry

Robert A Hoffman and David W Houck

11 Cell Separation Using Flow Cytometric Cell Sorting

Part VI: Special Applications

Gregor Siebenkotten, Katja Petry, Ute Behrens-Jung, Stefan Miltenyi, and Andreas Radbruch

12 Employing Surface Markers for the Selection of Transfected Cells

Hassan T Hassan, K Gutensohn, A R Zander, and P Kuehnl

13 CD34+ Cell Sorting and Enrichment: Applications in Blood Banking and Transplantation

Page 325

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

Avidin, columns, 11 (table), 88–89, 89 (figure)

Page 326mononuclear cells of, 63, 65, 66 (table), 67, 68 (figure), 74 (figure)

Catcher tube sorting, 248–249, 248 (figure)

Cell cycle, separation of cells according to, 51, 258

direct versus indirect, 183–186, 184 (figure), 185 (figure)

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

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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

Page 327Centrifugation, 24, 25, 27, 28 (figure), 52

CFU, see Colony-forming units

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)

Dendritic cells, 51, 53, 82, 103, 104, 126, 162, 163 (figure)

Density-adjusted cell sorting (DACS), 29–34

Density gradient material,

characteristics of, 17

Epithelial cells, 51, 225, 226 (figure)

Erythrocytes, 2 3 50, 115, 117, 162, 164 (figure), 223, 224 (table), 264lysis of, 2

Erythroid burst-forming cells (E-BFC), 70

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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)

Granulocyte-macrophage colony-forming cell (GM-CFC), 70, 95, 95 (table), 96 (table)

GVHD, see Graft-versus-host disease

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, 275continuous, 211–233

Investigational device exemption (IDE), 140

Investigational new drug (IND), 140

Iron, 155

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acute lymphocytic (ALL), 81, 286

acute myelogenous (AML), 81, 286

Page 329Long-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, 279Lymphoma, 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

MHC, see Major histocompatiblity class

Monocytes, 1 18, 43, 50, 51 (figure), 53, 162, 163 (figures), 223

mpl ligand, 285, 290

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

PBMC, see Peripheral blood mononuclear cells

PBPC, see Peripheral blood progenitor cells

PEG, see Polyethylene glycol

Start of Citation[PU]Marcel Dekker, Inc.[/PU][DP]1998[/DP]End of Citation

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

Page 330Polymerase chain reaction (PCR), 106, 124, 125, 135, 145, 198, 249, 259

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, 289Recombinant human interleukin (rHuIL)

Red blood cell, see Erythrocytes

Renal cell carcinoma, 81