Cell and Tissue Reaction Engineering: With a Contribution by Martin Fussenegger and Wilfried Weber

364 Pages·2009·5.294 MB·English

by  Prof. Dr.-Ing. Regine Eibl, Prof. Dr.-Ing. Dieter Eibl, PD Dr.-Ing. Ralf Pörtner, Prof. Dr. Eng. Gerardo Catapano, Prof. Dr.-Ing. Peter Czermak (auth.)

Principles and Practice
Regine Eibl • Dieter Eibl • Ralf Pörtner
Gerardo Catapano • Peter Czermak
Cell and Tissue Reaction
Engineering
With a Contribution by Martin Fussenegger
and Wilfried Weber
Prof. Dr.-Ing. Regine Eibl Prof. Dr. Eng. Gerardo Catapano
Prof. Dr.-Ing. Dieter Eibl Department of Chemical Engineering
Institute for Biotechnology and Materials
Zurich University of Applied Sciences University of Calabria
Department for Life Sciences and Facility 87030 Rende (CS), Italy
Management e-mail: [email protected]
P.O. Box
8820 Wädenswil, Switzerland
e-mail: [email protected]
e-mail: [email protected]
PD Dr.-Ing. Ralf Pörtner Prof. Dr.-Ing. Peter Czermak
Hamburg University of Technology Institute of Biopharmaceutical Technology
(TUHH) University of Applied Sciences
Institute for Bioprocess and Biosystems Giessen-Friedberg
Engineering Wiesenstrasse 14
Denickestr. 15 35390 Giessen, Germany
21073 Hamburg, Germany and
e-mail: [email protected] Department of Chemical Engineering
Kansas State University, Manhattan, USA
e-mail: [email protected]
ISBN: 978-3-540-68175-5 e-ISBN: 978-3-540-68182-3
DOI: 10.1007/978-3-540-68182-3
Library of Congress Control Number: 2008926595
© 2009 Springer-Verlag Berlin Heidelberg
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Cover design: WMX Design GmbH, Heidelberg, Germany
Cover illustration: Alginate-encapsulated human mesenchymal stem cell line (hMSC-TERT) © Institute
of Biopharmaceutical Technology, University of Applied Sciences Giessen-Friedberg, Giessen, Germany
Printed on acid-free paper
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Preface
The completion of the Human Genome Project and the rapid progress in cell biol-
ogy and biochemical engineering, are major forces driving the steady increase of
approved biotech products, especially biopharmaceuticals, in the market. Today
mammalian cell products (“products from cells”), primarily monoclonals,
cytokines, recombinant glycoproteins, and, increasingly, vaccines, dominate the
biopharmaceutical industry. Moreover, a small number of products consisting of
in vitro cultivated cells (“cells as product”) for regenerative medicine have also
been introduced in the market.
Their efficient production requires comprehensive knowledge of biological as
well as biochemical mammalian cell culture fundamentals (e.g., cell characteristics
and metabolism, cell line establishment, culture medium optimization) and related
engineering principles (e.g., bioreactor design, process scale-up and optimization).
In addition, new developments focusing on cell line development, animal-free cul-
ture media, disposables and the implications of changing processes (multi-purpose-
facilities) have to be taken into account. While a number of excellent books treating
the basic methods and applications of mammalian cell culture technology have been
published, only little attention has been afforded to their engineering aspects.
The aim of this book is to make a contribution to closing this gap; it particularly
focuses on the interactions between biological and biochemical and engineering
principles in processes derived from cell cultures. It is not intended to give a com-
prehensive overview of the literature. This has been done extensively elsewhere.
Rather, it is intended to explain the basic characteristics of mammalian cells related
to cultivation systems and consequences on design and operation of bioreactor sys-
tems, so that the importance of cell culture technology, as well as differences com-
pared to microbial fermentation technology, is understood.
The book is divided into a part related to mammalian cells in general (part I) and
a second part discussing special applications (part II). Part I starts with an overview
of mammalian cell culture technology in Chapter 1, where we present definitions,
the historical perspective and application fields including products; we summarize
the most commonly used mammalian cell types as well as their characteristics and
discuss resulting process requirements in Chapter 2. This chapter also outlines the
current understanding of the cell metabolism including mammalian cell culture
kinetics and modelling. Chapters 3 to 5 cover engineering aspects of mammalian
v
vi Preface
cell culture technology: Chapter 3 includes a categorization of suitable mamma-
lian cell culture bioreactor types with their descriptions, and highlights the trends
for R&D and biomanufacturing, as well as special features of bioreactors for cell
therapy and tissue engineering. Chapters 4 and 5 discuss key process limitation
issues and show how they should be handled with respect to optimized mammalian
cell bioreactor and process design. In this context we distinguish between suspen-
sion cells (non-anchorage dependent growing cells) and adherent cells (anchorage
dependent growing cells) and focus on shear stress by aeration as well as mixing,
oxygen transport, nutrient transfer, process strategy and monitoring, scale-up fac-
tors and strategies.
In part II, special cell culture applications which are predicted to have a high
potential for further development are treated (see Chapter 6 to Chapter 8). These
chapters treat insect cell-based recombinant protein production, bioartificial organs
and plant cell-based bioprocessing.
The questions and problems included should enable the reader, especially stu-
dents, to develop a clear understanding of the fundamentals and interactions pre-
sented in each chapter of the book. We hope that this book will be of interest to
students of biotechnology who specialize in cell cultivation techniques and bio-
chemical engineering as well as to more experienced scientists and industrial users
who work with cell cultures as production organisms.
Finally, we acknowledge our contributing authors, Dr. Wilfried Weber and Prof.
Dr. Martin Fussenegger and thank them for their dedication and diligence. We
would also like to thank our families for their support during the writing and editing
of this book. In addition, we are very grateful to Springer for their keen interest in
bringing out this book of quality work.
May 2008 G. Catapano
P. Czermak
R. Eibl
D. Eibl
R. Pörtner
Contents
Part I Mammalian Cells
1 Mammalian Cell Culture Technology: An Emerging Field................. 3
D. Eibl, R. Eibl, and R. Pörtner
1.1 Definition and History ........................................................................ 3
1.2 Fields of Application and Products from Mammalian Cells .............. 6
1.3 Future Prospects ................................................................................. 8
1.4 Exercises ............................................................................................. 9
References ................................................................................................... 9
Complementary Reading ............................................................................. 11
2 Characteristics of Mammalian Cells
and Requirements for Cultivation .......................................................... 13
R. Pörtner
2.1 Differences Between Mammalian Cells, Plant Cells
and Microbes: Consequences of These Differences ............................. 13
2.2 Types of Mammalian Cells ............................................................... 14
2.2.1 From Primary Cells to Permanent
(Established) Cell Lines ........................................................ 15
2.2.2 Hybridom Cells for Production of Monoclonal
Antibodies ............................................................................. 17
2.2.3 Culture Collections and Cell Banking................................... 20
2.3 Culture Media ................................................................................... 21
2.4 Characteristics of Cell Growth and Metabolism ............................... 24
2.4.1 Short Introduction to Cell Metabolism ................................. 24
2.4.2 Glucose, Glutamine and Amino Acids as Carbon
and Energy Source ................................................................ 26
2.4.3 The Effects of Lactate and Ammonia ................................... 29
2.4.4 Oxygen Uptake and Carbon Dioxide Production .................. 31
2.5 Kinetic Modelling of Cell Growth and Metabolism ......................... 32
2.5.1 Introduction to Kinetic Modelling for Mammalian Cells ..... 32
2.5.2 Set-Up of an Unstructured Model ......................................... 33
vii
viii Contents
2.5.3 Structured Models ................................................................. 44
2.5.4 Conclusions for Set-Up of a Kinetic Model .......................... 45
2.6 Questions and Problems .................................................................... 46
List of Symbols .......................................................................................... 47
References .................................................................................................. 47
Complementary Reading ........................................................................... 53
3 Bioreactors for Mammalian Cells: General Overview......................... 55
D. Eibl and R. Eibl
3.1 Technical Terminology: Bioreactor/Fermentor,
Bioreactor Facility ............................................................................. 56
3.2 Suitable Bioreactor Types for Mammalian Cell Cultures ................. 57
3.2.1 Categorization, Functional Principle, Possible
Fields of Application ............................................................. 57
3.2.2 Bioreactor Trends and the Increasing Acceptance
of Disposables ....................................................................... 66
3.3 Special Case: Bioreactors for Patient-Specific
Therapies Based on Functional Tissue and Stem Cells ..................... 69
3.3.1 Bioreactors for Growing 3D Tissues ..................................... 70
3.3.2 Bioreactors for Large-Scale Expansion
and Differentiation of Stem Cells......................................... 72
3.4 Conclusions ....................................................................................... 73
3.5 Questions and Problems .................................................................... 74
List of Abbreviations and Symbols ............................................................ 74
References .................................................................................................. 75
Complementary Reading ........................................................................... 82
4 Special Engineering Aspects ................................................................... 83
P. Czermak, R. Pörtner, and A. Brix
4.1 Cell Damage by Shear and Aeration ................................................. 83
4.1.1 General Aspects .................................................................... 83
4.1.2 Model Analysis ..................................................................... 85
4.1.3 Cell Damage in Bioreactors .................................................. 92
4.2 Oxygen Supply .................................................................................. 102
4.2.1 Introduction ........................................................................... 102
4.2.2 Limitations for Oxygen Transfer ........................................... 105
4.2.3 Oxygen Supply Systems (Aeration Systems) ....................... 108
4.2.4 Consequences for Reactor Design and Operation ................. 120
4.3 Immobilization of Cells .................................................................... 122
4.3.1 Carriers for Cell Immobilization ........................................... 123
4.3.2 Encapsulation ........................................................................ 134
Contents ix
4.4 Culture Modes ................................................................................... 136
4.4.1 Principles of Culture Modes.................................................. 136
4.4.2 Examples of Different Culture Modes .................................. 141
4.4.3 Process Strategies for Fed-Batch .......................................... 147
4.4.4 Process Strategies Applied in Industrial Processes ............... 149
4.5 Monitoring and Controlling in Animal Cell Culture ........................ 152
4.5.1 Temperature ........................................................................... 152
4.5.2 pH .......................................................................................... 153
4.5.3 Oxygen Partial Pressure ........................................................ 154
4.5.4 Carbon Dioxide Partial Pressure ........................................... 156
4.5.5 Metabolites and Products ...................................................... 156
4.5.6 Cell Density and Viability ..................................................... 159
4.5.7 Agitation ................................................................................ 160
4.6 Questions and Problems .................................................................... 160
4.6.1 Problem ................................................................................. 160
4.6.2 Problem ................................................................................. 160
List of Symbols .......................................................................................... 162
References .................................................................................................. 164
Complementary Reading ........................................................................... 171
5 Bioreactor Design and Scale-Up ............................................................. 173
G. Catapano, P. Czermak, R. Eibl, D. Eibl, and R. Pörtner
5.1 Bioreactor Design ............................................................................. 173
5.1.1 Bioreactors for Suspended Cells ........................................... 176
5.1.2 Fixed Bed and Fluidized Bed Bioreactors: Design,
Performance and Scale-Up .................................................... 199
5.1.3 Membrane Bioreactors .......................................................... 217
5.1.4 Disposable Bioreactors.......................................................... 242
5.2 Selection of Bioreactor and Operation Mode ................................... 245
5.3 How to Grow Mammalian Cells from Cryopreserved
Vial to Production Bioreactor ............................................................ 246
5.4 Questions and Problems .................................................................... 249
List of Symbols .......................................................................................... 250
References .................................................................................................. 253
Complementary Reading ........................................................................... 259
Part II Special Applications
6 Insect Cell-Based Recombinant Protein Production ............................ 263
W. Weber and M. Fussenegger
6.1 Insect Cell Culture ............................................................................ 263
6.2 Special Aspects: Engineering Baculoviruses as Vectors................... 264
6.3 Bioreactor Concepts .......................................................................... 267
x Contents
6.4 Process Design .................................................................................. 268
6.4.1 Optimization of Nutrient Supply ........................................... 268
6.4.2 Optimization of Production Kinetics .................................... 268
6.5 Applications ...................................................................................... 270
6.6 Current Trends in Insect Cell-Based Protein Production .................. 270
6.6.1 Elimination of Product Protein Proteolysis ........................... 273
6.7 Limitations ........................................................................................ 273
References .................................................................................................. 274
Complementary Reading ........................................................................... 276
7 Bioreactors for Bioartificial Organs ....................................................... 279
G. Catapano
7.1 Introduction ....................................................................................... 279
7.2 Cells for Bioartificial Organs ............................................................ 280
7.2.1 Expression of the Adult Cell Phenotype ............................... 283
7.3 Bioreactor Design for Bioartificial Organs ....................................... 285
7.3.1 Extravascular (EV) Bioreactors ............................................ 289
7.3.2 Intravascular (IV) Bioreactors ............................................... 295
7.3.3 Membrane Immunoprotection
in Bioartificial Organs ........................................................... 302
7.4 Commercial Bioreactors and Applications ....................................... 304
List of Symbols .......................................................................................... 310
References .................................................................................................. 311
8 Plant Cell-Based Bioprocessing .............................................................. 315
R. Eibl and D. Eibl
8.1 Plant Cell Culture Basics .................................................................. 316
8.1.1 Characteristics of Plant Cells and Culture Conditions .......... 316
8.1.2 Media..................................................................................... 317
8.1.3 Plant Cell Culture Types and Their In Vitro Initiation .......... 319
8.1.4 Routine Working Methods in Plant Cell Cultivation ............ 326
8.2 Bioreactors for Plant Cell Cultures ................................................... 330
8.2.1 General Considerations ......................................................... 330
8.2.2 Suitable Bioreactors for Plant Cell Suspension Cultures ...... 332
8.2.3 Suitable Bioreactors for Hairy Roots .................................... 335
8.3 Approaches to Improving Productivity in Plant Cell-Based
Bioprocessing .................................................................................... 340
8.4 Application Examples and Potential Active
Agent Candidates .............................................................................. 342
8.5 Conclusions ....................................................................................... 343
8.6 Questions and Problems .................................................................... 344
Contents xi
List of Abbreviations and Symbols ............................................................ 345
References .................................................................................................. 347
Complementary Reading ........................................................................... 356
Index ............................................................................................................... 357