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E-raamat: Animal Cell Culture and Technology

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(University of Manitoba, Canada)
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Butler (University of Manitoba) describes the basic requirements for establishing and maintaining cell cultures both in the laboratory and in large-scale operations. For this second edition, new topics are included to reflect the latest developments and trends in the field, covering the latest theory of the biological clocks (telomeres) of cell lines, the development of serum-free media formulations, the increased understanding and control of protein glycosylation, and the humanization of antibodies for therapeutic use. Annotation ©2004 Book News, Inc., Portland, OR (booknews.com)

Animal cell culture is an important laboratory technique in the biological and medical sciences. It has become an essential tool for the study of most biochemical and physiological processes and the use of large-scale animal cell culture has become increasingly important to the commercial production of specific compounds for the pharmaceutical industry. This book describes the basic requirements for establishing and maintaining cell cultures both in the laboratory and in large-scale operations. Minimal background knowledge of the subject is assumed and therefore it will be a readable introduction to animal cell culture for undergraduates, graduates and experienced researchers. Reflecting the latest developments and trends in the field, the new topics include the latest theory of the biological clock of cell lines, the development of improved serum-free media formulations, the increased understanding of the importance and control of protein glycosylation, and the humanization of antibodies for therapeutic use.
Abbreviations ix
Preface xi
Chapter 1 Introduction: The use of animal cell culture 1(10)
1. Tissue culture, organ culture and cell culture
1(1)
2. Why grow animal cells in culture?
1(1)
3. The advantages and disadvantages of using cell culture
2(1)
4. The early history of cell culture
3(2)
5. How long will primary cultures survive?
5(1)
6. The biological clock
6(1)
7. The first products of animal cell technology
7(4)
Chapter 2 Characteristics of cells in culture 11(16)
1. Where to obtain cells?
11(1)
2. Cells from tissue: a primary culture
11(2)
3. Cell types
13(1)
4. How to select a particular cell type
14(1)
5. What is a 'normal' cell?
15(1)
6. Anchorage-dependence
16(1)
7. The culture of differentiated cells
17(1)
8. Embryonic stem cells
18(1)
9. Adult stem cells
19(1)
10. Transformed cells
20(1)
11. Cells from a culture collection
21(3)
Protocol 2.1: Preparing a primary cell culture of chick embryo fibroblasts
24(2)
Protocol 2.2: Isolation of lymphocytes from a blood sample
26(1)
Chapter 3 Basic equipment and laboratory design: what you need to get started in cell culture 27(20)
1. Introduction
27(1)
2. Laboratory design
27(2)
3. Washing re-usable glassware
29(1)
4. Biosafety/laminar flow cabinets
30(1)
5. Incubators
31(3)
6. Laboratory-scale culture flasks
34(7)
7. Microscopes
41(1)
8. Centrifugation
42(1)
9. Liquid nitrogen storage
42(3)
10. Osmometer
45(2)
Chapter 4 Growth and maintenance of cells in culture 47(20)
1. How to culture cells and what to expect
47(4)
2. The importance of aseptic techniques
51(3)
3. Culture conditions
54(1)
4. Culture medium
55(3)
5. Media supplements
58(2)
6. Cell metabolism during culture
60(4)
Protocol 4.1: Harvesting anchorage-dependent cells with trypsin
64(1)
Protocol 4.2: Adaptation of cells to a serum-free medium
65(1)
Protocol 4.3: The trypsinization of cells in serum-free medium
66(1)
Chapter 5 Cell line and culture monitoring 67(32)
1. Cell counting and monitoring
67(3)
2. Indirect methods of cell determination
70(2)
3. Viability measurements
72(2)
4. Cell line identification
74(4)
5. Analysis of the cell cycle
78(5)
Protocol 5.1: Determination of the concentration of viable cells in a suspension by hemocytometer counting
83(1)
Protocol 5.2: Determination of the nuclei concentration in a culture sample
84(1)
Protocol 5.3: Determination of cell concentration by a Coulter counter
85(1)
Protocol 5.4: Determination of the protein concentration of a cell suspension
86(1)
Protocol 5.5: Determination of the DNA concentration of a cell suspension
87(2)
Protocol 5.6: Determination of the glucose concentration in culture
89(3)
Protocol 5.7: Determination of the viable concentration of cells by dye exclusion: the tetrazolium assay
92(1)
Protocol 5.8: Determination of cell viability by lactate dehydrogenase determination
93(1)
Protocol 5.9: Determination of the intracellular energy charge
94(2)
Protocol 5.10: Determination of the cellular rate of protein synthesis
96(1)
Protocol 5.11: Cell characterization by Giemsa banding of chromosomes (karyotype analysis)
97(2)
Chapter 6 Genetic engineering of animal cells in culture 99(14)
1. Introduction
99(1)
2. The need for mammalian cells
100(1)
3. How to get DNA into mammalian cells
100(3)
4. How to select and amplify the genes of transfected cells
103(2)
5. How to get genes to express proteins
105(2)
6. Regulation of gene expression
107(2)
Protocol 6.1: Transfection of CHO-K1 cells by lipofection
109(1)
Protocol 6.2: Transfection by electroporation
110(1)
Protocol 6.3: Development and isolation of methotrexate-resistant CHU cells
111(2)
Chapter 7 The glycosylation of proteins in cell culture 113(22)
1. Introduction
113(1)
2. Glycan structures present in glycoproteins
114(1)
3. Assembly and processing of glycans on proteins
115(4)
4. Glycoprotein analysis
119(1)
5. Why mammalian cells are chosen for glycoprotein production
120(2)
6. Control of glycan processing in mammalian cell culture
122(5)
7. Genetic engineering of mammalian cells to modify glycosylation
127(3)
8. Conclusion
130(3)
Protocol 7.1: Analysis of glycans from in-gel release of protein bands
133(2)
Chapter 8 Hybridomas - sources of antibodies 135(20)
1. Introduction
135(1)
2. Antibody production in vivo
135(1)
3. The structure of antibodies
136(1)
4. Glycosylation of antibodies
137(1)
5. Monoclonal antibodies
138(1)
6. Hybridomas
138(7)
7. Assay of monoclonal antibodies
145(2)
8. Human monoclonal antibodies
147(1)
9. Recombinant antibodies
148(3)
10. Potential alternative methods of production
151(1)
11. The importance of glycosylation to therapeutic antibodies
152(1)
12. Conclusion
153(2)
Chapter 9 Scaling up animal cell culture 155(20)
1. Why scale-up cultures?
155(1)
2. The stirred tank reactor (STR)
155(10)
3. Process control
165(1)
4. Alternative types of bioreactors
166(9)
Chapter 10 Modes of culture for high cell densities 175(20)
1. Batch culture
175(4)
2. Fed-batch culture
179(1)
3. Continuous culture
180(7)
4. Cell immobilization
187(6)
Protocol 10.1: Establishment and monitoring of a microcarrier culture
193(2)
Chapter 11 Production from cell culture 195(10)
1. Why use animal cell culture for production?
195(1)
2. How to produce biologicals from cell culture
195(4)
3. How to purify the final product
199(1)
4. The efficiency and productivity of a culture system
200(3)
5. Cost of the cell culture process
203(2)
Chapter 12 Mammalian cell products: established and potential 205(28)
1. Introduction
205(1)
2. Viral vaccines
205(4)
3. Monoclonal antibodies
209(2)
4. Therapeutic recombinant glycoproteins from mammalian cells
211(10)
5. Risks associated with cell culture products
221(2)
6. Tissue engineering
223(1)
7. Cells as products
224(3)
8. Conclusion
227(6)
Glossary 233(8)
Index 241
Michael Butler
Lisainfo e-raamatute kohta Lisainfo e-raamatute kohta
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