10 Insecta (Blattaria)
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1. Cut an antenna.
Bleeding
2. Collect the blood that oozes out.
1. Place the blood collected into a culture vessel with culture medium. Culture
2. Keep the culture at 26°C.
set-up
Comments
The culture may be improved by the use of conditioned medium. To prepare conditioned medium, S-20 medium with 20% FBS has been used
for culturing the embryonic cell line EPa, derived from P. americana, for
48 h (Landureau and Grellet, 1975). The medium was filtered with a membrane filter (0.22 flm; Millipore),and the filtrate was mixed with an equal
volume of fresh S-20 medium.
Results
Cells survive for several months if the medium that is used has been
supplemented with 20% FBS treated at 60°C for 1 h. Cells may multiply,
and subculture will be possible. Using this method, a continuous cell
line has been established from P. americana (Landureau and Grellet,
1975).
I
Subprotocol 3
Nervous Tissues
The following technique is based on the method of Chen and LeviMontalcini (1970).
Materials
- Dissecting microscope
Equipment
- Small glass tubes (5-10 ml capacity)
- Maximov slides
- Small glass container (13 mm diameter, 7 mm height; Fig. 2)
- Cover slips
- Pasteur pipettes
- Petri dishes
- Culture medium (a mixture of 5 parts Schneider's Drosophila medium Solutions
and 4 parts Eagle's basal amino acid solution for vertebrate cells, with
1 mg/ml streptomycin and 100 IV/ml penicillin)
- Newly laid oothecae of P. americana
Animals