260
Organ Culture
Results
Using this method, Laverdure (1975) cultured ovaries of the meal worm,
and observed that ovarian and muscular tissue extracts ensure the young
oocytes survival by trophic action. Laverdure (1975) also confirmed that
vitello genic protein in the culture medium was taken up by the oocytes
without intervention of the corpora allata hormone. In the case of lepidopteran insects, ovaries have been cultured mostly to obtain migrated
cells from ovarian epithelium. When the ovaries of lepidopteran insects
are cultured as a whole, the ovaries usually do not show marked changes
during the culture.
Subprotocol 17
Ovaries and Fat Bodies of the Honeybee (Hymenoptera)
Materials
Equipment - Dissecting microscope and dissecting tray (see chapter I, section 3
"Tools")
- Scissors
- Forceps
- Needles
- Pasteur pipettes
- Culture vessel (round glass dish)
Solutions - Honeybee medium (see Appendix 1)
Animals - Adult worker bees or the queen of Apis mellifera
Preparation
of adult bees
Dissection
Culture
set-up
Procedure
1. Collect honeybees 7 or 8 days after emergence from the brood nest of
colonies.
1. Isolate the abdomen, and pull away the last two segments to remove
the sting apparatus, gut, and Malpighian tubules.
2. Cut the wall of the abdominal body laterally along the pleural membrane into ventral and dorsal halves in physiological solution placed
in a dissecting tray, and discard the former.
3. For culturing fat bodies, use the dorsal body wall as an explant, because the fat body is attached to the body wall as a single layer, and it
cannot be dissected out without being damaged.
4. For culturing ovaries, excise the ovary from the body wall preparation of an egg-laying queen.
1. Place the excised organ in a round glass dish containing 150 f.11 honeybee medium so that the epithelium of the explants floats on the sur-
Organ Culture
Results
Using this method, Laverdure (1975) cultured ovaries of the meal worm,
and observed that ovarian and muscular tissue extracts ensure the young
oocytes survival by trophic action. Laverdure (1975) also confirmed that
vitello genic protein in the culture medium was taken up by the oocytes
without intervention of the corpora allata hormone. In the case of lepidopteran insects, ovaries have been cultured mostly to obtain migrated
cells from ovarian epithelium. When the ovaries of lepidopteran insects
are cultured as a whole, the ovaries usually do not show marked changes
during the culture.
Subprotocol 17
Ovaries and Fat Bodies of the Honeybee (Hymenoptera)
Materials
Equipment - Dissecting microscope and dissecting tray (see chapter I, section 3
"Tools")
- Scissors
- Forceps
- Needles
- Pasteur pipettes
- Culture vessel (round glass dish)
Solutions - Honeybee medium (see Appendix 1)
Animals - Adult worker bees or the queen of Apis mellifera
Preparation
of adult bees
Dissection
Culture
set-up
Procedure
1. Collect honeybees 7 or 8 days after emergence from the brood nest of
colonies.
1. Isolate the abdomen, and pull away the last two segments to remove
the sting apparatus, gut, and Malpighian tubules.
2. Cut the wall of the abdominal body laterally along the pleural membrane into ventral and dorsal halves in physiological solution placed
in a dissecting tray, and discard the former.
3. For culturing fat bodies, use the dorsal body wall as an explant, because the fat body is attached to the body wall as a single layer, and it
cannot be dissected out without being damaged.
4. For culturing ovaries, excise the ovary from the body wall preparation of an egg-laying queen.
1. Place the excised organ in a round glass dish containing 150 f.11 honeybee medium so that the epithelium of the explants floats on the sur-