Cryopreservation in Transgenic Suspension Cell Cultures of Oryza sativa L.

초록

Transgenic suspension cells of Oryza sativa L. utilized as a host for producing recombinant proteins were preserved in liquid nitrogen (-196℃) after slow pre-freezing in deep freezer (-70℃). The development of optimal procedure for long-term storage was investigated by adding various concentrations of cryoprotectant mixture and osmoticum in pre-culture media before cooling. Pre-freezing time was effective at 120 min on cell viability. 0.5 M sucrose compared with mannitol, sorbitol, trehalose, NaCl on the same osmotic condition was found to be the most effective osmoticum for pre-culture media. The cryoprotectant comprising sucrose, glycerol and dimethylsulfoxide (DMSO) was applied to the pre-cultured cells and combination of 1 M sucrose, 1 M glycerol, 1 M DMSO provided the best result for Oryza sativa L. This condition gave 88% survival rate of Oryza sativa L. after immersion into liquid nitrogen.