Effects of Amino Acids on Cell Viability and hCTLA4Ig Production in Transgenic Rice Cell Cultures

초록

Rice cells were transformed to produce human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig). The target protein was produced and secreted when sugar was depleted in culture medium by transgenic rice cell lines (Oryza sativa) using RAmy3D promoter. To induce RAmy3D promoter system, problems such as increase of cell death and release of protease due to sugar starvation should be overcome. In this study, we evaluate the effects of amino acids on cell viability and hCTLA4Ig production in transgenic rice cell cultures. Functions of amino acids (arginine, aspartic acid, glutamine, glycine), as a nitrogen and carbon source were examined to enhance cell viability and the production of hCTLA4Ig. In addition, selected 10 amino acids were examined at various concentrations of amino acids (1∼2 mM) were supplemented in induction phase. A Plackett–Burman statistical design was conducted to minimize the number of experiments. It was found that long-term survival of transgenic rice cells and enhanced production of hCTLA4Ig were possible by feeding a mixture of amino acids with optimized concentrations in transgenic rice cell cultures.