21 Insecta
263
Procedure
1. Select third instar larvae (approximately 96 h after hatching).
2. Wash the larvae with sterilized distilled water.
1. Dissect the larvae in Ringer's solution placed in the depression of a
Maximov slide under a dissecting microscope. There are two types of
melanotic tumor tissues; one floats in the hemolymph and the other
is embedded in various tissues, such as fat bodies, hypodermis, intestine, and trachea.
2. Excise the melanotic tumors that are embedded in tissues.
3. Wash the tumors in Ringer's solution.
1. Transfer the tumors into a culture vessel with culture medium.
2. Culture at 25°C,
Results
According to Kuroda and Tamura (1955), when the tumors floating in
hemolymph were cultured, no growth of tumorous tissues was observed,
whereas when tumors embedded in tissues were cultured, marked melanotic growth was observed in the surrounding tissues. However, this
melanotic growth seemed to be due to the melanosis of the tissues surrounding the tumors, and not due to the multiplication of tumorous cells.
The melanotic growth of the tissues was markedly inhibited by co culture
with cephalic complex containing ring glands.
Subprotocol 20
Dechorionated Egg Culture and Further Development up to
Imaginal Stage in the Silkworm (Lepidoptera)
Materials
- Dissecting microscope
- Forceps
- Needles
- Petri dishes (4-5 em diameter)
Preparation
of larvae
Dissection
Culture
set-up
Equipment
- Physiological solution
Solutions
- Solution A (NaCI 670 mg, KCI 30 mg, CaCI 2 ·2H 2 0 20 mg,
MgS0 4·7H 2 0 20 mg, glucose 50 mg, lactalbumin hydrolysate
300 mg/lOO ml distilled water)
- Mix 80 ml Solution A with 10 ml of 1/15 M KH 2 P0 4 and 10 ml of 1/
15 M Na 2 HP0 4 ·12H 2 0)
- Grace's culture medium without serum or hemolymph and pH adjusted to 7.5 (see Appendix 1)
263
Procedure
1. Select third instar larvae (approximately 96 h after hatching).
2. Wash the larvae with sterilized distilled water.
1. Dissect the larvae in Ringer's solution placed in the depression of a
Maximov slide under a dissecting microscope. There are two types of
melanotic tumor tissues; one floats in the hemolymph and the other
is embedded in various tissues, such as fat bodies, hypodermis, intestine, and trachea.
2. Excise the melanotic tumors that are embedded in tissues.
3. Wash the tumors in Ringer's solution.
1. Transfer the tumors into a culture vessel with culture medium.
2. Culture at 25°C,
Results
According to Kuroda and Tamura (1955), when the tumors floating in
hemolymph were cultured, no growth of tumorous tissues was observed,
whereas when tumors embedded in tissues were cultured, marked melanotic growth was observed in the surrounding tissues. However, this
melanotic growth seemed to be due to the melanosis of the tissues surrounding the tumors, and not due to the multiplication of tumorous cells.
The melanotic growth of the tissues was markedly inhibited by co culture
with cephalic complex containing ring glands.
Subprotocol 20
Dechorionated Egg Culture and Further Development up to
Imaginal Stage in the Silkworm (Lepidoptera)
Materials
- Dissecting microscope
- Forceps
- Needles
- Petri dishes (4-5 em diameter)
Preparation
of larvae
Dissection
Culture
set-up
Equipment
- Physiological solution
Solutions
- Solution A (NaCI 670 mg, KCI 30 mg, CaCI 2 ·2H 2 0 20 mg,
MgS0 4·7H 2 0 20 mg, glucose 50 mg, lactalbumin hydrolysate
300 mg/lOO ml distilled water)
- Mix 80 ml Solution A with 10 ml of 1/15 M KH 2 P0 4 and 10 ml of 1/
15 M Na 2 HP0 4 ·12H 2 0)
- Grace's culture medium without serum or hemolymph and pH adjusted to 7.5 (see Appendix 1)