250
Organ Culture
- Scissors
- Needle
- Pasteur pipettes
- Cover glasses
- Culture dishes (internal diameter 13 mm, height 7 mm)
Solutions - Insect Ringer's solution (see Appendix 1)
- Culture medium (a mixture of 5 parts Schneider's insect solution and
4 parts Eagle's basal medium, containing penicillin 100 IU Iml, streptomycin 100 jlg/ml, and fungizone 125 jlg/ml)
Animals - Nymphs (2-6 months old) and adults (within 1 week after adult molt)
of the American cockroach P. americana
Procedure
Sterilization 1. Anesthetize the animals with CO 2 •
2. Fix the animal, dorsal side up, to a Maximov slide using Scotch tape.
3. Expose the neck by applying light pressure to the scotch tape across
the thorax with the head of the cockroach tucked inward.
4. Sterilize the dorsal cervical sclerites using absorbent cotton moistened with 95% ethanol.
Dissection 1. Make a slit in this region, and excise the corpora cardiaca and allata
together using fine forceps.
2. Transfer the excised complex to sterile insect Ringer's solution.
3. Remove any adhering tissues, and wash the complex several times in
insect Ringer's solution.
Culture 1. Transfer the complex into culture medium.
set-up 2. Press the complex gently against small cover glasses (10 x 10 mm) in
order to stick the organs to the glass.
3. Place the cover glasses with the organ complex in culture dishes containing 0.25 ml culture medium.
Results
Seshan and Levi-Montalcini (1971) cultured the corpora cardiaca-allata
complex either alone or in combination with brain, pharynx, esophagus,
and heart dissected from embryos of the same species. A large number
of cells migrated out from the explants of corpora cardiaca. Fewer cells
migrated from the corpora allata. These migrating cells were identified
as glial or interstitial cells on the basis of their features. A few cells with
characteristics of neurons were seen in the migratory zone around explants of corpora cardiaca. The cells and fibers from these explants formed
connections between themselves and neighboring embryonic explants.
Holbrook et al. (1997) cultured corpora allata of the cockroach Diploptera
Organ Culture
- Scissors
- Needle
- Pasteur pipettes
- Cover glasses
- Culture dishes (internal diameter 13 mm, height 7 mm)
Solutions - Insect Ringer's solution (see Appendix 1)
- Culture medium (a mixture of 5 parts Schneider's insect solution and
4 parts Eagle's basal medium, containing penicillin 100 IU Iml, streptomycin 100 jlg/ml, and fungizone 125 jlg/ml)
Animals - Nymphs (2-6 months old) and adults (within 1 week after adult molt)
of the American cockroach P. americana
Procedure
Sterilization 1. Anesthetize the animals with CO 2 •
2. Fix the animal, dorsal side up, to a Maximov slide using Scotch tape.
3. Expose the neck by applying light pressure to the scotch tape across
the thorax with the head of the cockroach tucked inward.
4. Sterilize the dorsal cervical sclerites using absorbent cotton moistened with 95% ethanol.
Dissection 1. Make a slit in this region, and excise the corpora cardiaca and allata
together using fine forceps.
2. Transfer the excised complex to sterile insect Ringer's solution.
3. Remove any adhering tissues, and wash the complex several times in
insect Ringer's solution.
Culture 1. Transfer the complex into culture medium.
set-up 2. Press the complex gently against small cover glasses (10 x 10 mm) in
order to stick the organs to the glass.
3. Place the cover glasses with the organ complex in culture dishes containing 0.25 ml culture medium.
Results
Seshan and Levi-Montalcini (1971) cultured the corpora cardiaca-allata
complex either alone or in combination with brain, pharynx, esophagus,
and heart dissected from embryos of the same species. A large number
of cells migrated out from the explants of corpora cardiaca. Fewer cells
migrated from the corpora allata. These migrating cells were identified
as glial or interstitial cells on the basis of their features. A few cells with
characteristics of neurons were seen in the migratory zone around explants of corpora cardiaca. The cells and fibers from these explants formed
connections between themselves and neighboring embryonic explants.
Holbrook et al. (1997) cultured corpora allata of the cockroach Diploptera