[转移帖]分享细胞微载体培养的好书
原帖由论坛会员qhg发表于 2011-2-14 14:41 |:Contents
1 Introduction .......................................................................................................... 7
1.1 Reasons for using microcarriers .............................................................................. 9
1.1.1 New opportunities and applications for animal cell culture ..................................................................10
1.1.2 Increased production capacity ..........................................................................................................10
1.1.3 Separating cells from secreted product .............................................................................................10
1.1.4 Improved control .............................................................................................................................11
1.1.5 Protection against physical and chemical stress .................................................................................11
1.1.6 Reduced requirements for culture medium ........................................................................................12
1.1.7 Reduced requirements for labor .......................................................................................................12
1.1.8 Lower risk of contamination .............................................................................................................12
1.2 Aim of this book .................................................................................................. 12
2 Microcarrier background .................................................................................... 13
2.1 Adhesion (cell-cell, cell-surface) ........................................................................... 13
2.1.1 Adhesion to microcarriers ................................................................................................................17
2.2 Immobilization principles ..................................................................................... 19
2.3 Materials ............................................................................................................ 20
2.4 Size, shape and diffusion limits ............................................................................ 21
2.5 Specific density and sedimentation velocity ........................................................... 22
2.6 Rigidity and shear force ....................................................................................... 22
2.7 Porosity .............................................................................................................. 22
2.8 Cell observation ................................................................................................... 23
3 Microcarrier technology ...................................................................................... 25
3.1 The development of microcarriers for animal cell culture .......................................................................25
3.2 Advantages of microcarriers .................................................................................. 27
3.3 Possible drawbacks ............................................................................................. 28
3.4 Scale-up considerations ....................................................................................... 28
3.5 Choice of supplier ............................................................................................... 29
4 Applications ....................................................................................................... 31
4.1 Vaccines ............................................................................................................. 31
4.1.1 Production of viruses and vectors .....................................................................................................32
4.2 Natural and recombinant proteins ......................................................................... 34
4.2.1 Comparison of microcarriers in different reactors (packed bed or fluidized bed reactor) ..........................35
4.2.2 Long-term production with perfused macroporous microcarriers ...........................................................35
4.2.3 Process stability of perfusion with macroporous microcarriers ..............................................................36
4.3 Monoclonal antibodies ......................................................................................... 37
4.3.1 Comparison of anti-HIV monoclonal antibody productivity with different processes ................................38
4.3.2 Comparison of a hollow fiber reactor with a fluidized bed reactor .........................................................38
4.4 Potential future applications ................................................................................. 39
4.5 Further applications ............................................................................................. 40
4.5.1 Production of large numbers of cells .................................................................................................40
4.5.2 Studies on cell function, metabolism and differentiation .....................................................................41
4.5.3 Proteolytic enzyme-free subcultivation and cell transfer ......................................................................43
4.5.4 Microscopy ....................................................................................................................................44
4.5.5 Harvesting mitotic cells ...................................................................................................................53
4.6 Which microcarrier to use? ................................................................................... 54
4.6.1 Decision tree .................................................................................................................................54
4.6.2 Further comments on selecting Cytodex microcarriers .........................................................................58
5 Microcarrier products ......................................................................................... 61
5.1 Cytodex microcarriers ........................................................................................... 61
5.1.1 Cytodex 1 ......................................................................................................................................61
5.1.2 Cytodex 3 ......................................................................................................................................62
5.1.3 Cell types cultured on Cytodex microcarriers ......................................................................................64
5.2 Cytopore microcarriers ......................................................................................... 67
5.2.1 Cell types cultured on Cytopore ........................................................................................................68
5.3 Cytoline microcarriers .......................................................................................... 69
5.3.1 Cell types cultured on Cytoline .........................................................................................................71
6 Microcarrier culture equipment ........................................................................... 73
6.1 General outline of procedure ................................................................................. 73
6.2 Requirements ..................................................................................................... 74
6.2.1 Siliconizing culture vessels ..............................................................................................................74
6.3 Unit process system ............................................................................................ 74
6.4 Small-scale equipment ........................................................................................ 75
6.4.1 Spinner and rod-stirred vessels .........................................................................................................75
6.4.2 Roller bottles .................................................................................................................................77
6.4.3 Rocking bottles shaker flasks ...........................................................................................................79
6.4.4 Air-lift and fluid-lift culture systems ..................................................................................................79
6.4.5 Small-scale perfusion culture ...........................................................................................................80
6.4.6 Dishes, tubes and multi-well trays ....................................................................................................80
6.4.7 Bench-scale fermenters ...................................................................................................................80
6.5 Large-scale equipment (culture vessels) ................................................................. 81
6.5.1 Large-scale stirred tank microcarrier culture vessels ...........................................................................81
6.5.2 Packed beds .................................................................................................................................82
6.5.3 Fluidized beds ...............................................................................................................................83
6.5.3.1 Fluidized bed with external circulation ................................................................................................................. 83
6.5.3.2 Cytopilot – fluidized bed with internal circulation .................................................................................................. 83
6.5.3.3 Fluidization and fluidization velocity .................................................................................................................... 84
6.6 Retention systems ............................................................................................... 85
6.6.1 Spin filters ....................................................................................................................................85
6.6.2 Settlers .........................................................................................................................................86
6.6.3 Ultrasonic device ............................................................................................................................86
6.6.4 Hollow fibers .................................................................................................................................87
6.6.5 Centrifuges ...................................................................................................................................88
7 Culture conditions ............................................................................................... 89
7.1 Media and components ........................................................................................ 89
7.1.1 Choice of culture medium ................................................................................................................89
7.1.1.1 General comments on components of culture media .............................................................................................. 90
7.1.1.2 Practical aspects of culture media ....................................................................................................................... 91
7.1.1.3 Amino acids ...................................................................................................................................................... 91
7.1.1.4 Nucleic acid precursors ..................................................................................................................................... 92
7.1.1.5 Carbon sources and lactate ................................................................................................................................ 93
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