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KMID : 1094720000050050372
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Biotechnology and Bioprocess Engineering
2000 Volume.5 No. 5 p.372 ~ p.378
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Development of cell line preservation method for research and industry producing useful metabolites by plant cell culture
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Cho Ji-Suk
Chun Su-Hwan
Lee Song-Jae
Kim Ik-Hwan
Kim Dong-Il
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Abstract
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The cell culture ofAngelica gigas Nakai producing decursin derivatives and immunostimulating polysaccharides was preserved in liquid nitrogen after pre-freezing in a deep freezer at ?70°C for 480 min. The effects of the cryoprotectant and pretreatment before cooling were investigated to obtain the optimal procedure for cyropreservation. When compared to mannitol, sorbitol, or NaCl with a similar osmotic pressure, 0.7M sucrose was found to be the best osmoticum for the cryopreservation ofA. gigias cells. In the pre-culture medium, the cells in the exponential growth phase showed the best post-freezing survival after cryopre-servation. A mixture of sucrose, glycerol, and DMSO was found to be an effective cryoprotectant and a higher concentration of the cryoprotectant provided better cell viability. When compared with the vitrification, the optimum cryopreservation method proposed in this study would seem to be more effective for the long-term storage of suspension cells. The highest relative cell viability established with the optimal procedure was 89%.
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KEYWORD
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Angelica gigas Nakai, cryopreservation, vitrification, cryoprotectant
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