Flow Cytometry Applications in Cell Culture

344 Pages·1996·79.922 MB·English

by  Mohamed Al-Rubeai, A Nichol Emery

Flow Cytometry
Applications in
Cell Culture
edited by
Mohamed M-Rubeai
A. Nicholas Emery
School of Chemical Engineering
The University of Birmingham
Birmingham, England
Marcel Dekker, Inc. New York.Basel.Hong Kong
Library of Congress Cataloging-in-Publication Data
Flow cytometry applications in cell cultureI edited by Mohamed AI-Rubeai, A. Nicholas
Emery.
cm p..
Includes index.
ISBN: 0-8247-9614-4 (hbk. :a lk. paper)
1. Flow cytometry. 2. Cell culturIe.. Al-Rubeai, Mohamed,I I. Emery, A. Nicholas.
QH5855F56F5714 9 96
514.87’0724-dc20 95-40326
CIP
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PRINTED IN THE UNITED STATES OF AMERICA
Preface
Flow cytometry is now a standard technique, with many applications in the
biological and medical sciences, and much effort has been applied to advancing
the instrumentation and techniques in both research and diagnostic fields. More
recently this has led to growing interest in its use for the identification, charac-
terization, monitoring, and control of microbial, plant, and animal cell biopro-
cesses. It is now possible to obtain information about cell populations unobtain-
able by other means and of a form and quality far superior to that available by
conventional biochemical analysis and that can be available in a near on-line
mode and timescale. With such information readily available, it should be possi-
ble to make significant progress in the optimization of large-scale cell culture for
the successful manufacture of those many valuable products that are arising as a
direct result of recent developments in molecular and cell biology, and even to
consider the flowc ytometera s a near on-linep rocess sensor for control
purposes. In particular, the increasing availability of fluorescing markers for the
analysis of cell structure and function is likely to make significant contributions
to the elucidation of the population dynamics and hence the development of
sophisticated models of cell growth and products ynthesis and secretion.
This book brings together the knowledge and experience of those who are
applyingf l owc ytometryt o the investigationa ndm onitoringo f cellular
processes for biotechnological purposes. Chapters review the state of the art with
in-depth assessments that emphasize the practical aspects of efficient operation
of flow cytometrict echniques. The book indicates those aspects of cell variabil-
ity encountered in cell culture processes and shows howt he quantitative analysis
of heterogeneous populationsc an help full realizationo f the potential of
biotechnological exploitation of cells. Cell cycle analysis of animal cell cultures
is of particular importance as an aid to the early estimation of process state and
to the prediction of future culture performance. Many of the authors lay empha-
sis on such studies. The importance of cell line selection and product stability
are now widely recognized, and informationi s presented here on the use of flow
cytometry to monitora nd improve the selection and enrichment ofh igh-produc-
tivity cell lines. Analysis ofb aculovirus-infected insect cell culture is dealt with
iii
iV Preface
as are a number of flow cytometry techniques of varying complexity that have
been developed to detect cellular and subcellelular changes occurring during
apoptosis.
mow cytometry offers a rapid and versatile means to analyze the proper-
ties of hematopoietic cells. Its use to characterize ex vivo expandedh uman bone
marrow is presented with detailed protocols. The applications of flow cytometry
to studies of microbial populations are also described, including the determina-
tion of population structure and cell cycle progression in yeast and the analysis
of bacterial growth and viability. The potential for flow cytometry to character-
ize plant cell populations together with the problems associated with analysis
and sorting of plant cells are included to provide a sound basis for future devel-
opments in this newly opened area.
Individual chapters provide reviews by experienced practitioners of the
specific methodologies available together with outlines of recent data obtained
and insights gained. The material is addressed primarily to those interested in
flow cytometry in microbial and cell culture processes, but many topics will also
be of interest to microbiologists, biochemists, and other bioscientists as well as
to biochemical engineers. The editors come from bioscience (Al-Rubeai) and
engineering (Emery) backgrounds and are therefore well placed to ensure that
the contents are addressed to and understandable by a wide range of disciplines.
We are enthusiasts for the cause of flow cytometry-we hope that our readers
find inspiration to explore further its potential themselves and that this work
helps their rapid progress.
Mohamed Al-Rubeai
A. NicholaEs mery
'.
Contents
Preface iii
Contributors ix
Monitoring the Proliferative Capacity of Cultured Animal
Cells by Cell Cycle Analysis 1
V. Leelavatcharamas, A. Nicholas Emery,a nd
Mohumed Al-Rubeai
Use of Now Cytometry for Monitoring Antibody Productivity
and Isolating High-Secreting Hybridoma Cells 17
Katherine L. McKinney and Georges Belfort
Antibody Secretion Assays Using Gel Microdrops and
Flow Cytometry 39
James C. Weaver, JohnF . Dunne, Forest Gray,
Kristina G. Lauari, and James B. Lin
High-Resolution Cell Cycle Analysis of Cell Cycle-
Regulated Gene Expression 63
Manfred Kubbies, Bernhard Goller, and Guenter Giese
Stability of Monoclonal Antibody Production in Hybridoma
Cell Culture 85
Josb M. Coco Martina nd E. Coen Beuvery
Flow Cytometric Studies of Osmotically Stressed and
Sodium Butyrate-Treated Hybridoma Cells 101
Steve Oh, FlorenceK . F. Chua, and Mohamed Al-Rubeai
V
vi Contents
7 Flow Cytometric Analysis of Cells Obtained from
Human Bone Marrow Cultures 121
David Brott, Manfred R. Koller, Sue A. Rummel,
and Bernhard Palsson
8 Measurement of Intracellular pH During the Cultivation
of Hybridoma Cells in Batch and Continuous Cultures 147
Jean-Marc Engasser, Annie Marc, Marc Cherlet,
Pierre. Nabeta, nd Patricia Franck
9 The Relationship Between Intracellular pH and Cell Cycle
in Cultured Animal Cells Using SNARF-1 Indicator 163
Jonathan P. Welsh and Mohamed Al-Rubeai
10 Assessment of Cell Viability in Mammalian Cell Lines 177
Xavier Ronot, Sylvain Paillasson, and KatherAi.n Me uirhead
11 Analysis of Apoptosis Cbyy tFo mlowetr y 193
Anne E. Milner, Hong Wang, and Christopher D. Gregory
12 Monitoring Baculovirus Infection and Protein Expression
in Insect Cells 21 1
Anthanassia K. Kioukia, N. H. Simpson, Jatin D. Shah,
A. Nicholas Emerya, nd Mohamed Al-Rubeai
13 Use of Protein Distribution to Analyze Budding Yeast
Population Structure and Cell Cycle Progression 225
Danilo Porro and Lilia Alberghina
14 Cell Cycle Analysis of Microorganisms 241
Kirsten Skarstad,R olfBernander, Sture Wold,
Harald B. Steen, and Erik Boye
15 Bacterial Characterization by Flow Cytometry 257
Gerhard Nebe-von Caron and R. Andrew Badley
Contents vi1
16 Assessmentof Viability of Bacteria by Flow Cytometry 29 l
Clive Edwards
17 Advances in the Flow Cytometric Characterization of
Plant Cells and Tissues 311
David W.G albraith and Georgina M. Lambert
Index 327
This Page Intentionally Left Blank
Contributors
MohamedA l-Rubeai School ofC hemical Engineering,T heU niversity of
Birmingham, Birmingham, England
Lilia Alberghina Dipartimento di Fisiologia e Biochimica Generali, Sezione
Biochimica Comparata, Universith degli Studi di Milano, Milan,I taly
R. Andrew Badley Immunology Department, Colworth Laboratory, Unilever
Research, Sharnbrook, Bedford, England
Georges Belfort Department of Chemical Engineering, Rensselaer Polytechnic
Institute, Troy, New York
Rolf Bernander Departmento fB iophysics, Institute for CancerR esearch,
Oslo, Norway
E. Coen Beuvery National Institute for PublicH ealth andE nvironmental
Protection (RIVM), Bilthoven, The Netherlands
Erik Boye Department of Biophysics, Institute for CancerR esearch, Oslo,
Norway
David Brott Aastrom Biosciences, Inc., Ann Arbor, Michigan
Marc Cherlet Laboratoire des Sciences du Gdnie Chimique-CNRS, Institut
National Polytechnique deL orraine, Vandceuvre-15s-Nancy,F rance
Florence K. F. Chua Bioprocessing Technology Centre, National University
of Singapore, Singapore
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