256
Organ Culture
- Pasteur pipettes
- Maximov slides
- Culture vessels
Solutions - Physiological solution
- Culture media (Grace's medium or a modification of Grace's medium;
see Appendix 1)
Animals - Female adults of the Indian meal moth Plodia interpunctella at various ages
Procedure
Sterilization 1. Surface sterilize the moths by submersion in 70% ethanol for 10 min.
2. Wash the moths with sterilized distilled water.
Dissection 1. Remove the wings of the moths.
2. Fix the moths on a dissecting tray filled with physiological solution.
3. Isolate the sex pheromone gland (SPG), which is comprised of a single
layer of specially differentiated epidermal cells with surrounding tissues.
4. Remove most of the attached tissues, such as the gut, oviduct and fat
bodies.
Culture 1. Rinse the SPG in physiological solution.
set-up 2. Transfer the gland into a culture vessel with 1 ml culture medium.
Results
According to Asokasrinivasan et al. (1978), the SPG cells appeared normal, even after 10 days culture in either chemically defined Grace's medium or a modification of this medium (modified by the addition of whole
egg ultrafiltrate, FBS and bovine serum albumin fraction V). The sex
pheromone (Z-E)-9,12-tetradecadien-l-01 acetate was recovered from the
cultured medium.
Subprotocol 14
Proventriculus of the Blowfly (Diptera)
Materials
Equipment - Dissecting microscope
- Fine scissors
- Forceps
- Needles
- Petri dishes
Organ Culture
- Pasteur pipettes
- Maximov slides
- Culture vessels
Solutions - Physiological solution
- Culture media (Grace's medium or a modification of Grace's medium;
see Appendix 1)
Animals - Female adults of the Indian meal moth Plodia interpunctella at various ages
Procedure
Sterilization 1. Surface sterilize the moths by submersion in 70% ethanol for 10 min.
2. Wash the moths with sterilized distilled water.
Dissection 1. Remove the wings of the moths.
2. Fix the moths on a dissecting tray filled with physiological solution.
3. Isolate the sex pheromone gland (SPG), which is comprised of a single
layer of specially differentiated epidermal cells with surrounding tissues.
4. Remove most of the attached tissues, such as the gut, oviduct and fat
bodies.
Culture 1. Rinse the SPG in physiological solution.
set-up 2. Transfer the gland into a culture vessel with 1 ml culture medium.
Results
According to Asokasrinivasan et al. (1978), the SPG cells appeared normal, even after 10 days culture in either chemically defined Grace's medium or a modification of this medium (modified by the addition of whole
egg ultrafiltrate, FBS and bovine serum albumin fraction V). The sex
pheromone (Z-E)-9,12-tetradecadien-l-01 acetate was recovered from the
cultured medium.
Subprotocol 14
Proventriculus of the Blowfly (Diptera)
Materials
Equipment - Dissecting microscope
- Fine scissors
- Forceps
- Needles
- Petri dishes