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Methods for Setting Up Primary Cultures Specific to Animal Groups
I Subprotocol 6
Wing Discs
Wing discs are located inside the stigmas of meso- and metathoraces.
They exist only in the larval stage. The·discs develop markedly during
the last larval stage. This development must be associated with cell proliferation. Thus, many attempts have been made to obtain proliferating
cells from these discs.
Materials
Equipment The equipment used is the same as that given for subprotocol1.
Solutions - Physiological solution
- Culture media
Animals - The last instar larvae of lepidopteran insects
Procedure
Sterilization Follow the instructions given for sterilization in sub protocol 1.
Dissection 1. Follow steps 1-4 for dissection given in subprotocol 1.
2. Remove the wing discs. The wing discs are located near the first and
second stigmas (Fig. 7), and are somewhat difficult to find because
they are located between the muscles and cuticle, and are surrounded
by many small fat body lobes. However, the wing discs look glittering
whitish -silver due to condensed tracheation. Pluck out the small whitish mass inside the lateral cuticle of the meso- and metathoraces together with the surrounding tissue.
3. Remove extra tissues. Ring-shaped hemocytopoietic organs (cf. chapter 5, Fig. 14) attach firmly to.the wing discs. Separate both organs.
Culture 1. Cut the discs into several pieces.
set-up 2. Wash the fragments in physiological solution, and transfer them into
culture medium.
3. Transfer the fragments, with the culture medium, into culture vessels.
4. Culture at 25°C.
Results
Cell migration will occur from the cut opening of the wing discs. Sometimes migrated cells multiply by forming spherical vesicles. If the vesicles
increase in size and number, the culture may be subcultured as a sus-
Methods for Setting Up Primary Cultures Specific to Animal Groups
I Subprotocol 6
Wing Discs
Wing discs are located inside the stigmas of meso- and metathoraces.
They exist only in the larval stage. The·discs develop markedly during
the last larval stage. This development must be associated with cell proliferation. Thus, many attempts have been made to obtain proliferating
cells from these discs.
Materials
Equipment The equipment used is the same as that given for subprotocol1.
Solutions - Physiological solution
- Culture media
Animals - The last instar larvae of lepidopteran insects
Procedure
Sterilization Follow the instructions given for sterilization in sub protocol 1.
Dissection 1. Follow steps 1-4 for dissection given in subprotocol 1.
2. Remove the wing discs. The wing discs are located near the first and
second stigmas (Fig. 7), and are somewhat difficult to find because
they are located between the muscles and cuticle, and are surrounded
by many small fat body lobes. However, the wing discs look glittering
whitish -silver due to condensed tracheation. Pluck out the small whitish mass inside the lateral cuticle of the meso- and metathoraces together with the surrounding tissue.
3. Remove extra tissues. Ring-shaped hemocytopoietic organs (cf. chapter 5, Fig. 14) attach firmly to.the wing discs. Separate both organs.
Culture 1. Cut the discs into several pieces.
set-up 2. Wash the fragments in physiological solution, and transfer them into
culture medium.
3. Transfer the fragments, with the culture medium, into culture vessels.
4. Culture at 25°C.
Results
Cell migration will occur from the cut opening of the wing discs. Sometimes migrated cells multiply by forming spherical vesicles. If the vesicles
increase in size and number, the culture may be subcultured as a sus-