Chapter 9
Insecta (Hymenoptera)
Many attempts have been made to culture bee cells without success. However, several cell lines have been established from the sawfly (Sohi and
Ennis, 1981) and from endoparasitoid wasps (Lynn and Hung, 1986, 1991).
The culture media used for hymenopteran cell cultures are modified
Grace's medium and modified Goodwin's IPL-S2B (see Appendix 1).
Subprotocol 1
Ovaries
Ovaries of queen bees, taken from larval as well as adult stages of the
honeybee Apis mellifera, have been cultured. The following procedures
are based on the methods of Giauffret et al. (1968).
Materials
PROTOCOL
- Dissecting microscope
Equipment
- Dissecting tray (see chapter I, section 3 "Tools")
- Pasteur pipettes
- Scissors
- Forceps
- Needles
- Small petri dishes
- Physiological solution
Solutions
- Culture medium
- Either larvae of honeybees that are destined to become queen or queen Animals
honeybees
Procedure
1. Surface sterilize the insects by submersion in 70% ethanol for 10 min. Sterilization
1. Fix the insects in a dissecting tray.
Dissection
2. Fill the tray with appropriate physiological solution.
3. Make an incision in the abdomen, and remove the ovaries.
4. Remove any attached connective tissue.
5. Transfer the ovaries into culture medium placed in a small petri dish.