240
Organ Culture
Results
Agui and Fukaya (1973) cultured wing discs of the cabbage armyworm
Mamestra brassicae in Grace's medium (see Appendix 1), and observed
that the development of the discs was significantly accelerated by the
addition of ecdysteroids. Oberlander and Leach (1979) cultivated the wing
discs of the greater wax moth Galleria mellonella and the Indian meal
moth Plodia interpunctella in Grace's medium. During culture, migration of the tracheoles into the lacunae of the discs, evagination of the
discs, and the production of pupal cuticle were observed, and these processes were stimulated in the presence of 20-hydroxyecdysone and inhibited by juvenile hormone.
Subprotocol 2
Imaginal Discs of Flies (Diptera)
With regard to dipteran insects, many attempts have been made to culture imaginal discs of fruit flies for studies on the differentiation of tissues. The techniques adopted in these studies are similar, and the following is based on the technique of Horikawa (1959).
Materials
Equipment - Dissecting microscope
- Petri dishes
- Maximov slides
- Forceps
- Needles
- Pasteur pipettes
- Culture vessels
Solutions - Carlson's solution (see Appendix 1)
- Culture medium
Animals - Aseptically reared third ins tar larvae of the fruit fly Drosophila
melanogaster (see Comments)
Preparation
of larvae
Dissection
Procedure
1. Collect aseptically reared third instar larvae.
2. Wash the larvae in sterilized distilled water.
1. Transfer the larvae into the depression of a Maximov slide filled with
Carlson's solution.
2. Dissect the larvae using fine forceps and needles, and isolate imaginal
Organ Culture
Results
Agui and Fukaya (1973) cultured wing discs of the cabbage armyworm
Mamestra brassicae in Grace's medium (see Appendix 1), and observed
that the development of the discs was significantly accelerated by the
addition of ecdysteroids. Oberlander and Leach (1979) cultivated the wing
discs of the greater wax moth Galleria mellonella and the Indian meal
moth Plodia interpunctella in Grace's medium. During culture, migration of the tracheoles into the lacunae of the discs, evagination of the
discs, and the production of pupal cuticle were observed, and these processes were stimulated in the presence of 20-hydroxyecdysone and inhibited by juvenile hormone.
Subprotocol 2
Imaginal Discs of Flies (Diptera)
With regard to dipteran insects, many attempts have been made to culture imaginal discs of fruit flies for studies on the differentiation of tissues. The techniques adopted in these studies are similar, and the following is based on the technique of Horikawa (1959).
Materials
Equipment - Dissecting microscope
- Petri dishes
- Maximov slides
- Forceps
- Needles
- Pasteur pipettes
- Culture vessels
Solutions - Carlson's solution (see Appendix 1)
- Culture medium
Animals - Aseptically reared third ins tar larvae of the fruit fly Drosophila
melanogaster (see Comments)
Preparation
of larvae
Dissection
Procedure
1. Collect aseptically reared third instar larvae.
2. Wash the larvae in sterilized distilled water.
1. Transfer the larvae into the depression of a Maximov slide filled with
Carlson's solution.
2. Dissect the larvae using fine forceps and needles, and isolate imaginal