15 Mollusca
167
Fig. 5. Cell sheet consisting of
epithelial cell-like cells in the
culture of the mantle tissue of
Pinctada Jucata. Line indicates
200 11m. Courtesy of Dr. A.
Machii, Wakasa-Otsuki Pearl
Laboratory, Kashikojima, Japan
Fig. 6. Crystals formed on the explanted mantle tissue of Pinctada
Jucata (polarizing microscope). x 60.
From Machii, A. and Wada T. Some
marine invertebrates tissue culture, in
Mitsuhashi, J. (ed) "Invertebrate Cell
System Applications" Vol.I!. Fig. 3c.
Copyright © 1989 by CRC Press, Boca
Raton. Reprinted with permission of
copyright owner
- Wash the epithelia by dispase digestion with gentle pipetting in balanced salt solution containing 1 mg/ml collagenase (Sigma), 4 mg/ml
DNAse I (Sigma), and 0.4 mg/ml testicular hyaluronidase (Sigma)
- Centrifuge the suspension at 130 g for 5 min
- Suspend the sediment in culture medium
• • Results
Cell migration from the explant is initiated soon after the culture is set
up. In the case of culturing mantle tissue fragments of the pearl oyster
Pinctada fucata and the black lip pearl oyster Pinctada margaritifera,
Machii and Wada (1989) reported that numerous epithelial cell-like cells
and fibroblast-like cells appeared from the explants, and formed cell
sheets (Fig. 5). These authors also observed trefoil-like depositions at
the margin of the explants 35 days after the culture initiation (Fig. 6).
The trefoil-like depositions resemble prismatic crystals formed in vivo
at the initiation of pearl formation and shell regeneration.
Subprotocol 4
Foot and Mantle Tissues of Land Slugs (Pulmonata)
The following is the procedure used by Furuta (1989) to initiate a primary culture of the foot tissues of the land slug Incilaria frukstorferi.
167
Fig. 5. Cell sheet consisting of
epithelial cell-like cells in the
culture of the mantle tissue of
Pinctada Jucata. Line indicates
200 11m. Courtesy of Dr. A.
Machii, Wakasa-Otsuki Pearl
Laboratory, Kashikojima, Japan
Fig. 6. Crystals formed on the explanted mantle tissue of Pinctada
Jucata (polarizing microscope). x 60.
From Machii, A. and Wada T. Some
marine invertebrates tissue culture, in
Mitsuhashi, J. (ed) "Invertebrate Cell
System Applications" Vol.I!. Fig. 3c.
Copyright © 1989 by CRC Press, Boca
Raton. Reprinted with permission of
copyright owner
- Wash the epithelia by dispase digestion with gentle pipetting in balanced salt solution containing 1 mg/ml collagenase (Sigma), 4 mg/ml
DNAse I (Sigma), and 0.4 mg/ml testicular hyaluronidase (Sigma)
- Centrifuge the suspension at 130 g for 5 min
- Suspend the sediment in culture medium
• • Results
Cell migration from the explant is initiated soon after the culture is set
up. In the case of culturing mantle tissue fragments of the pearl oyster
Pinctada fucata and the black lip pearl oyster Pinctada margaritifera,
Machii and Wada (1989) reported that numerous epithelial cell-like cells
and fibroblast-like cells appeared from the explants, and formed cell
sheets (Fig. 5). These authors also observed trefoil-like depositions at
the margin of the explants 35 days after the culture initiation (Fig. 6).
The trefoil-like depositions resemble prismatic crystals formed in vivo
at the initiation of pearl formation and shell regeneration.
Subprotocol 4
Foot and Mantle Tissues of Land Slugs (Pulmonata)
The following is the procedure used by Furuta (1989) to initiate a primary culture of the foot tissues of the land slug Incilaria frukstorferi.