64
Methods for Setting Up Primary Cultures Specific to Animal Groups
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Fig. 17. Cells migrating from Malpighian tubes of larvae of the rice stem
borer Chilo suppressalis. x 100
Animals - Larvae of lepidopteran insects
Sterilization
and
dissection
Culture
set-up
• • Procedure
1. Follow the instructions given for sterilization, and steps 1-4 for dissection as listed in subprotocol 1.
2. Take the Malpighian tubes out.
1. Wash the tissues in physiological solution.
2. Transfer the tissues to culture medium.
3. Cut the tube into fragments less than 1 mm long.
4. Transfer the fragments, with the culture medium, into culture vessels.
5. Culture at 25°C,
• • Results
Cell migration occurs, but proliferation of the migrated cells is not expected if the present culture method is used (Fig. 17). It is necessary to
improve the culture conditions to get growing cells from Malpighian
tubes.
Subprotocol 11
Nervous Tissues
Usually, the central nervous system (CNS), starting from the brain and
ending at the last abdominal ganglion (Fig. 18), is cultured .
• • Materials
The materials are the same as those given in subprotocol 1.