The gametophytic cells of six species of Laminariales, Laminaria japonica Areschoug, L. longissima Miyabe, Kjellmaniella crassifolia Miyabe, Ecklonia stolonifera Okamura, E. kurome Okamura, and Undaria pinnatifida (Harvey) Suringar, were subjected to cryopreservation in liquid nitrogen. The cells were suspended in various cryoprotective solutions and slowly cooled to -401 C over a period of 4 h. After this slow cooling step, the suspensions were immediately immersed in liquid nitrogen. All the species of Laminariaceae used in the present study survived maximally in a mixture of ethylene glycol and proline. On the other hand, the gametophytic cells of Undaria pinnatifida, a member of the Alariaceae, survived maximally in the mixture of glycerol and proline. The viability of the thawed gametophytic cells decreased during postthawing incubation. The decrease in viability continued for 4-6 days, and the minimum levels ranged from 36.2% to 67.2%. After 4-6 days of incubation, the percentage viability of all strains began to increase due to the renewal of cell division.
Cryopreservation of sporothalli of the red alga Porphyra (P. seriata, P. yezoensis, P. tenera, P. pseudolinearis and P. dentata) was performed by the two-step cooling method in liquid nitrogen. The algal samples were suspended in various cryoprotective solutions, and slowly cooled to -40°C in 4 hours using a programmed freezer. After the first slow cooling the suspensions with cryoprotectants were immediately immersed in liquid nitrogen. The suspension from the programmed freezer was thawed quickly by agitation of the vial in a water bath at 40°C. When ice in the suspension of cryogenic vial was mostly melted, the vial was transferred to an ice bath for complete melting of the residual ice. The cryoprotectants in the vial were washed off by gradual dilution with seawater. The viability of the cell was assessed with neutral red staining. The viability of Porphyra samples ranged 54.6-70.9% when the mixed suspension of 10% dimethylsulfoxide and 0.5 M sorbitol in 50% seawater used as a cryoprotectant.
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