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Recently, production of recombinant proteins has been studied using transgenic plant cell cultures. Transgenic rice cell cultures (Oryza sativa L.) were utilized to produce recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig). General long-term preservation method for plant cells has been known to be the repeated subcultures. One drawback of this method is the genetic instability of transformed cell lines.
For that reason, a cryopreservation method was developed for the long-term storage of the transgenic rice cells. DMSO has been known to be an effective cryoprotectant in freezing steps, but plant cells are damaged by cryoprotectants during thawing procedures. To solve this problem, an optimal method has been designed by using various cryoprotectant mixtures in freezing step and the cryoprotectants were removed by negative pressure in thawing steps.
Optimization of cryoprotectant concentration was performed with range from 0 to 2 M. The best combination was found to be 1 M sucrose, 0.5 M glycerol and 0.5 M DMSO. The cell viability was 57.3% in that combination after 7days of cryopreservation. To remove toxic cryoprotectants in thawing steps, the times required for negative pressure treatment were maintained by 30 sec, 1 min, 2 min and 5 min, respectively. The best treatment time was found to be 2 min and cell viability was 84.7%. In conclusion, we could enhance the cell viability in cryopreservation of transgenic rice cells by optimizing cryoprotectants-related steps.

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저자

• Ji-Won CHOI [ Dept, of Biological Engineering, Inha University, Incheon 402-751. ]
• Jun-Young KWON [ Dept, of Biological Engineering, Inha University, Incheon 402-751. ]
• Su-Hwan CHEON [ Dept, of Biological Engineering, Inha University, Incheon 402-751. ]
• Yong-Suk YANG [ Dept, of Biological Engineering, Inha University, Incheon 402-751. ]
• Mi-Hee YOO [ Dept, of Biological Engineering, Inha University, Incheon 402-751. ]
• Soo-Jin KIM [ Dept, of Biological Engineering, Inha University, Incheon 402-751. ]
• Hyung-Jin NAM [ Dept, of Biological Engineering, Inha University, Incheon 402-751. ]
• Dong-Il KIM [ Dept, of Biological Engineering, Inha University, Incheon 402-751. ]

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간행물

표지보기 관심저널 등록

• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576