194
Methods for Setting Up Primary Cultures Specific to Animal Groups
Procedure
Dissection 1. Remove the white bodies from young animals (40-60 g) as soon as
the octopuses have been caught (Fig. 26).
Dissociation 1. Dissociate the tissues by pipetting in a small amount of octopus physiological solution.
2. Leave the tissue suspension for 2 min until the connective tissues sink.
Culture 3. Distribute the cell suspension into culture tubes containing 1 ml culset-up
ture medium.
4. Culture at 18°C by rotating the culture tubes in a rotation drum.
Results
According to Necco and Martin (1963), all cells attach well to the culture
tubes within a few minutes (Fig. 26). Mitosis can be observed at the beginning of culture.
References
Auzoux, S., Domart-Coulon, 1. and Doumenc, D. (1993) Gill cell cultures of the butterfish clam,
Ruditapes decussatus. J. Mar. Biotechnol. 1,79-81.
Awaji, M. (1991) Separation of outer epithelial cells from pearl oyster mantle by dispase digestion.
In: Franser, M. J. Jr. (ed.) Proceedings of the 8th International Conference on Invertebrate and
Fish Tissue Culture, June 16-20, 1991, Anaheim. Tissue Culture Association, Columbia, MD.,
pp 43-46.
Awaji, M. (1997) Primary culture techniques for the outer epithelial cells of pearl oyster mantle. In:
Maramorosch, K. and Mitsuhashi, J. (eds) Invertebrate Cell Culture: Novel Directions and
Biotechnology Applications. Science Publishers, Enfield, pp 239-244.
Awaji, M. and Suzuki, T. (1998) Monolayer formation and DNA synthesis of the outer epithelial
cells from pearl oyster mantle in co-culture with amebocytes. In Vitro Cell. Dev. BioI. 34 A,
486-491.
Boulo, V. and Cadoret,J.P. (1996) Transient expression of luciferase reporter gene after lipofection
in oyster (Crassostrea gigas) primary cell cultures. Mol. Mar. BioI. Biotechnol. 5, 167-174.
Burch, J.B. and Cuadros, C. (1965) A culture medium for snail cells and tissues. Nature 206, 637-638.
Cecil, J.T. (1969) Mitoses in cell cultures from cardiac tissue of the surf clam, Spisula solidissima. J.
Invert. Pathol. 14,407-410.
Chen, S.N., Jong, K.J. and Kou, G.H. (1989) Cell cultures derived from tissues of Penaeid shrimp,
Penaeus penicilatus, and hard clam, Meretrix lusoria. In: Mitsuhashi, J. (ed) Invertebrate Cell
System Applications, Vol. II. CRC, Boca Raton, pp 253-262.
Chernin, E. (1963) Observations on hearts explanted in vitro from the snail Australorbis glabratus.
J. Parasitol. 49, 353-364.
Dadkhah-Teherani, Z. and Bride, M. (1990-1991) Dissociation of the snail ovotestis: characterization and primary culture of isolated gonadal cells. BioI. Struct. Morphogen. 3, 71-82.
Domart-Coulon, 1., Oumenc,D., Auzoux-Bordenave, S. and Le Fichant, Y. (1994) Identification of
media supplements that improve the viability of primary cell cultures of Crassostrea gigas
oysters. Cytotechnology 16, 109-120.
Ebstein, B.S., Rosenthal, M.D. and Dehaan, R.L. (1965) Cells from isolated blastomeres of Ilyanassa
obsoleta in tissue culture. Exp. Cell Res. 40,174-177.
Methods for Setting Up Primary Cultures Specific to Animal Groups
Procedure
Dissection 1. Remove the white bodies from young animals (40-60 g) as soon as
the octopuses have been caught (Fig. 26).
Dissociation 1. Dissociate the tissues by pipetting in a small amount of octopus physiological solution.
2. Leave the tissue suspension for 2 min until the connective tissues sink.
Culture 3. Distribute the cell suspension into culture tubes containing 1 ml culset-up
ture medium.
4. Culture at 18°C by rotating the culture tubes in a rotation drum.
Results
According to Necco and Martin (1963), all cells attach well to the culture
tubes within a few minutes (Fig. 26). Mitosis can be observed at the beginning of culture.
References
Auzoux, S., Domart-Coulon, 1. and Doumenc, D. (1993) Gill cell cultures of the butterfish clam,
Ruditapes decussatus. J. Mar. Biotechnol. 1,79-81.
Awaji, M. (1991) Separation of outer epithelial cells from pearl oyster mantle by dispase digestion.
In: Franser, M. J. Jr. (ed.) Proceedings of the 8th International Conference on Invertebrate and
Fish Tissue Culture, June 16-20, 1991, Anaheim. Tissue Culture Association, Columbia, MD.,
pp 43-46.
Awaji, M. (1997) Primary culture techniques for the outer epithelial cells of pearl oyster mantle. In:
Maramorosch, K. and Mitsuhashi, J. (eds) Invertebrate Cell Culture: Novel Directions and
Biotechnology Applications. Science Publishers, Enfield, pp 239-244.
Awaji, M. and Suzuki, T. (1998) Monolayer formation and DNA synthesis of the outer epithelial
cells from pearl oyster mantle in co-culture with amebocytes. In Vitro Cell. Dev. BioI. 34 A,
486-491.
Boulo, V. and Cadoret,J.P. (1996) Transient expression of luciferase reporter gene after lipofection
in oyster (Crassostrea gigas) primary cell cultures. Mol. Mar. BioI. Biotechnol. 5, 167-174.
Burch, J.B. and Cuadros, C. (1965) A culture medium for snail cells and tissues. Nature 206, 637-638.
Cecil, J.T. (1969) Mitoses in cell cultures from cardiac tissue of the surf clam, Spisula solidissima. J.
Invert. Pathol. 14,407-410.
Chen, S.N., Jong, K.J. and Kou, G.H. (1989) Cell cultures derived from tissues of Penaeid shrimp,
Penaeus penicilatus, and hard clam, Meretrix lusoria. In: Mitsuhashi, J. (ed) Invertebrate Cell
System Applications, Vol. II. CRC, Boca Raton, pp 253-262.
Chernin, E. (1963) Observations on hearts explanted in vitro from the snail Australorbis glabratus.
J. Parasitol. 49, 353-364.
Dadkhah-Teherani, Z. and Bride, M. (1990-1991) Dissociation of the snail ovotestis: characterization and primary culture of isolated gonadal cells. BioI. Struct. Morphogen. 3, 71-82.
Domart-Coulon, 1., Oumenc,D., Auzoux-Bordenave, S. and Le Fichant, Y. (1994) Identification of
media supplements that improve the viability of primary cell cultures of Crassostrea gigas
oysters. Cytotechnology 16, 109-120.
Ebstein, B.S., Rosenthal, M.D. and Dehaan, R.L. (1965) Cells from isolated blastomeres of Ilyanassa
obsoleta in tissue culture. Exp. Cell Res. 40,174-177.