15 Mollusca
171
Fig. 10. Sterilization procedure of pearl oyster mantle. A An opened pearl oyster
(left), and after removal of a visceral mass (right). A pair of mantles (arrows) is left
on the shells. B Sterilization of the inner surface of the mantle. C The pallial zone of
a mantle (hatched) is cut out and used as material tissue; a, adductor muscle; m,
mantle; s, shell. D Rinsing of mantle fragments in balanced salt solution by pipetting.
From Awaji, M. (1997) with permission
2. Add 0.7 ml of the mixture of MMBSS and pearl oyster hemolymph
(2: 8), pH 7.3-7.4.
3. Culture at 25°C.
• • Results
The isolated cells are mostly epidermal cells with some mucous cells.
Very few dead cells or damaged cells will be observed at the beginning of
culture. Some cells will adhere to the substrate. The viability of the cells
after 3 days will be approximately 60%. During culture, there are indications that DNA synthesis occurs in migrated epithelial cells (Awaji and
Suzuki, 1998).
Subprotocol 6
Cardiac Tissues of Clams (Bivalvia)
• • Materials
- Centrifuge and SO-mi centrifuge tubes
- Surgical knife
- Stiff brush
- Forceps
- Beakers (SO ml)
- Petri dishes
- Magnetic stirrer and magnetic bar
- Culture vessels
- T-2s flasks
Equipment