19 Coelenterata
229
4. Centrifuge the cell suspension at 500 g for 20 min at 4°C.
5. Collect the red bands, and rinse the cells several times with SSW.
6. Disperse the final pellet into the culture medium, and dispense it into
wells of culture plates.
Comments
For dissociation of tissue, mechanical dissociation or chemical dissociation may be applied instead of spontaneous dissociation.
Results
Frank et al. (1994) cultured cells from 10 species of cnidarians, and obtained four continuously growing cell lines. The species from which the
cell lines were obtained were Stylophora pistillata (Anthoza), Clathrarcia
rubrinoides (Anthozoa),Plexaura A (Anthozoa) and Millepora dichotoma
(Hydrozoa). The cells from another six species proliferated also, and several passages were made, but these were frozen for future studies. For
dissociation of tissues, spontaneous dissociation was promising. This
technique has several advantages over other methods, including: (1) minimal cell damage and loss compared with other methods; (2) the capability to use this method for all species and all life stages; and (3) ease of
application. Frank et al. (1994) recommended the use of diluted culture
media for primary cultures. For example, 10% organic medium in seawater and 5%-10 % heat-inactivated FBS are the most suitable. The diluted medium improved the rate of cell adherence to the substrate. In
diluted medium, the cells tended to maintain the undifferentiated state.
When the cells were transferred to concentrated medium, the cell population became heterogeneous due to cell differentiation. The cell population obtained consisted mostly of scleroblasts, endodermal cells, ciliated
ectodermal cells, nermatoblasts, and undifferentiated interstitial cells.
References
Burnett, A.L., Ruffing, F.E., Zongker, J. and Necco, A. (1968) Growth and differentiation of Tubularia
cells in a chemically defined physiological medium. J. Embryo1. Exp. Morpho1. 20, 73-80.
Frank, u., Rabinowitz, C. and Rinkevich, B. (1994) In vitro establishment of continuous cell cultures
and cell lines from ten colonial cnidarians. Mar. BioI. 120,491-499.
Lewis, M.R. (1916) Sea water as a medium for tissue cultures. Anat. Rec. 10, 187-299.
Loomis, F.W. and Lenhoff, H.M. (1956) Growth and sexual differentiation of Hydra in mass culture.
J. Exp. Zoo1. 132,555-568.
Martin, R. and Tardent, P. (1963) Kultur von Hydroiden-Zellen in vitro. Rev. Suiss Zoo1. 70,312-316.
Rannou, M,. (1968) Formation de spicules dans des cultures cellulaires de cnidaire (Gorgone). Vie
Milieu Ser. A 19,53-57.
Youngner, J.S. (1954) Monolayer tissue cultures 1. Preparation and standardization of suspensions
of trypsin-dispersed monkey kidney cells. Proc. Soc. Exp. BioI. Med. 85, 202-205.
Yu-Ying, F.L., Baker, F.D. and Andrew, W. (1963) A method for tissue culture of Hydra cells. Proc.
Soc. Exp. BioI. Med. 113,259-262.
229
4. Centrifuge the cell suspension at 500 g for 20 min at 4°C.
5. Collect the red bands, and rinse the cells several times with SSW.
6. Disperse the final pellet into the culture medium, and dispense it into
wells of culture plates.
Comments
For dissociation of tissue, mechanical dissociation or chemical dissociation may be applied instead of spontaneous dissociation.
Results
Frank et al. (1994) cultured cells from 10 species of cnidarians, and obtained four continuously growing cell lines. The species from which the
cell lines were obtained were Stylophora pistillata (Anthoza), Clathrarcia
rubrinoides (Anthozoa),Plexaura A (Anthozoa) and Millepora dichotoma
(Hydrozoa). The cells from another six species proliferated also, and several passages were made, but these were frozen for future studies. For
dissociation of tissues, spontaneous dissociation was promising. This
technique has several advantages over other methods, including: (1) minimal cell damage and loss compared with other methods; (2) the capability to use this method for all species and all life stages; and (3) ease of
application. Frank et al. (1994) recommended the use of diluted culture
media for primary cultures. For example, 10% organic medium in seawater and 5%-10 % heat-inactivated FBS are the most suitable. The diluted medium improved the rate of cell adherence to the substrate. In
diluted medium, the cells tended to maintain the undifferentiated state.
When the cells were transferred to concentrated medium, the cell population became heterogeneous due to cell differentiation. The cell population obtained consisted mostly of scleroblasts, endodermal cells, ciliated
ectodermal cells, nermatoblasts, and undifferentiated interstitial cells.
References
Burnett, A.L., Ruffing, F.E., Zongker, J. and Necco, A. (1968) Growth and differentiation of Tubularia
cells in a chemically defined physiological medium. J. Embryo1. Exp. Morpho1. 20, 73-80.
Frank, u., Rabinowitz, C. and Rinkevich, B. (1994) In vitro establishment of continuous cell cultures
and cell lines from ten colonial cnidarians. Mar. BioI. 120,491-499.
Lewis, M.R. (1916) Sea water as a medium for tissue cultures. Anat. Rec. 10, 187-299.
Loomis, F.W. and Lenhoff, H.M. (1956) Growth and sexual differentiation of Hydra in mass culture.
J. Exp. Zoo1. 132,555-568.
Martin, R. and Tardent, P. (1963) Kultur von Hydroiden-Zellen in vitro. Rev. Suiss Zoo1. 70,312-316.
Rannou, M,. (1968) Formation de spicules dans des cultures cellulaires de cnidaire (Gorgone). Vie
Milieu Ser. A 19,53-57.
Youngner, J.S. (1954) Monolayer tissue cultures 1. Preparation and standardization of suspensions
of trypsin-dispersed monkey kidney cells. Proc. Soc. Exp. BioI. Med. 85, 202-205.
Yu-Ying, F.L., Baker, F.D. and Andrew, W. (1963) A method for tissue culture of Hydra cells. Proc.
Soc. Exp. BioI. Med. 113,259-262.