Development of Long-term Preservation Method for Transgenic Plant Cells Under Glucose-added Anoxia Condition

초록

Transgenic plant cell culture is an attractive alternative for the production of recombinant proteins such as pharmaceutical proteins due to the absence of contaminants associated with mammalian or other culture systems. Plant cells are routinely maintained by periodic subculture that is a major drawback of the stable maintenance of high-producing cell lines. To conform to existing regulatory requirements, plant cell cultures expressing biopharmaceuticals should be derived from a consistent and stable master stock. Cryopreservation would be an ideal method for retaining the original morphological forms and characteristics. However, cryopreservation of plant cells has not been established yet because of the enormous difficulties for cell banking such as osmotic pretreatment, cryoprotection, freezing, thawing and regeneration. It is known that rice is resistant to anoxia, and sugar availability plays an important role3). In this study, transgenic suspension cells of Oryza sativa L. utilized as a host for producing recombinant human cytotoxic T lymphocyte-associated antigen 4-immunoglobulin (hCTLA4Ig) were preserved in anoxia condition for 2 months. Cell viability was stably maintained at 76% for 2 months in the media containing 174 mM glucose under anoxia condition, and the cells were successfully recovered. There were no significant differences in the cell growth and hCTLA4Ig production between non-preserved and 2-month-preserved cells.