Basic studies of improved cryopreservation using supercooling technology.

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The cysts of Artemia franciscana (A. franciscana) were hydrated and their hatching rates were measured in order to estimate possible damagaes of cells and tissues by various freezing speeds. We observed that the hatching rates of hydrated cysts declined shortly at rapid frozen (submerged in liquid nitrogen), but high hatching rates maintained at slow freezing rate (0.5C/min), and that improved hatching ratio of decapsulated hydrated cysts by the treatment of cryoprotectants such as trehalose and dimethl sulfoxide (DMSO). We also evaluated that the surface of frozen hydrated cysts using a scanning electron microscope attached with cryo-transfer system (Cryo-SEM), and found fine structural discrepancy in the section of embryos between two freezing speeds.