Contents
Part 1 General Methods
Chapter 1 Facilities and Equipment ...................................................
3
1 Facilities .......................................................................................................
3
2 Equipment ...................................................................................................
3
3 Tools .............................................................................................................
10
4 Culture System ............................................................................................
13
Chapter 2 Basic Information and Overview...................................... 15
1 Routine Precautions for Sterile Work .......................................................
15
2 Culture Media ..............................................................................................
15
3 Primary Culture ..........................................................................................
22
Chapter 3 Preparation of Media .......................................................... 25
1 Direct Method of Preparing Media Consisting of a Few Chemicals ....
25
2 Indirect Method of Preparing Media Consisting of a Few Chemicals.
26
3 Preparation of Complex Media .................................................................
27
4 Preparation of Media Containing Water-Insoluble Substances ............
29
5 Sterilization of Culture Media ...................................................................
30
Chapter 4 General Cell Culture Methods ........................................... 33
1 Trypsin Treatment for Dissociation of Cell Masses ............................... 33
2 Pancreatin Treatment for Dissociation of Cell Masses .......................... 34
3 Dispase Treatment for Dissociation of Cell Masses ............................... 35
4 Methods for Subculturing ......................................................................... 36
5 Hybrid-Cell Technique for Obtaining Growing Cells ............................
37
6 Introduction of Oncogenes to Primary Cultured Cells to
Obtain Growing Cells ................................................................................. 39
7 Cryopreservation of Cells ..........................................................................
41
8 How to Get Cell Lines and How to Ship Cell Lines ................................
43
Part 2 Methods for Setting Up Primary Cultures Specific
to Animal Groups
Chapter 5 Insecta (Lepidoptera) ......................................................... 47
1 Ovaries ......................................................................................................... 47
2 Testes ............................................................................................................ 50
3 The Gut ........................................................................................................
51
4 Stem Cells from the Midgut ...................................................................... 52
5 Fat Bodies ..................................................................................................... 54
6 Wing Discs ...................................................................................................
56