5 Insecta (Lepidoptera)
51
cells may grow to a continuous cell lines (Mitsuhashi, 1965; Mano and
Mitsuhashi, 1981).
Subprotocol 3
The Gut
The gut of lepidopteran larvae consists of the foregut, midgut, and hindgut. To culture gut cells, insect material will preferably have been reared
aseptically. Otherwise, microorganisms in the gut will spoil the culture.
The insect should be starved for 24 h to empty the gut prior to dissection.
Materials
The materials to be used are the same as those given for subprotocol1. Equipment
- Physiological solution
Solutions
- Culture media
- Aseptically reared larvae of lepidopteran insects
Animals
Procedure
1. Follow the instructions given for sterilization, and steps 1-4 for dissection as listed in subprotocol 1.
2. Cut the gut at the anterior part of the foregut and at the hindgut.
3. Chop the gut into several pieces.
4. Wash the tissues in a physiological solution and transfer into culture
medium.
5. Transfer the fragments, with the culture medium, into culture vessels.
6. Culture at 25°C.
Results
Cell migration may occur soon after the culture is set up. However, migration and proliferation of cells is usually very slow, and the probability
of obtaining a continuously growing cell population is low. However, it is
possible to perform some experiments with the migrated cells in the primary culture. Generally, epithelioid-type cells migrate from gut explants
(Mitsuhashi, 1965; Mano and Mitsuhashi, 1981).
51
cells may grow to a continuous cell lines (Mitsuhashi, 1965; Mano and
Mitsuhashi, 1981).
Subprotocol 3
The Gut
The gut of lepidopteran larvae consists of the foregut, midgut, and hindgut. To culture gut cells, insect material will preferably have been reared
aseptically. Otherwise, microorganisms in the gut will spoil the culture.
The insect should be starved for 24 h to empty the gut prior to dissection.
Materials
The materials to be used are the same as those given for subprotocol1. Equipment
- Physiological solution
Solutions
- Culture media
- Aseptically reared larvae of lepidopteran insects
Animals
Procedure
1. Follow the instructions given for sterilization, and steps 1-4 for dissection as listed in subprotocol 1.
2. Cut the gut at the anterior part of the foregut and at the hindgut.
3. Chop the gut into several pieces.
4. Wash the tissues in a physiological solution and transfer into culture
medium.
5. Transfer the fragments, with the culture medium, into culture vessels.
6. Culture at 25°C.
Results
Cell migration may occur soon after the culture is set up. However, migration and proliferation of cells is usually very slow, and the probability
of obtaining a continuously growing cell population is low. However, it is
possible to perform some experiments with the migrated cells in the primary culture. Generally, epithelioid-type cells migrate from gut explants
(Mitsuhashi, 1965; Mano and Mitsuhashi, 1981).