Effect of various carbohydrates and amino acids on the cell viability and production of CTLA4Ig in Transgenic Rice Cell Culture

초록

Transgenic rice cells were transformed to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig). Target protein was secreted when sugar was depleted in culture medium by transgenic plant cell lines (Oryza sativa L.) using RAmy3D promoter. RAmy3D promoter system was induced cell death because the promoter is induced only by sugar starvation. In this study, enhancement of cell viability and hCTLA4Ig productivity using transgenic cell line was investigated in flask cultures by supplementing alternative energy sources such as carbohydrates and amino acid. Lactose and galactose were used as alternative sugar sources to active RAmy3D promotor and increase cell viability. supplementation of 30mM lactose and 60mM galactose in culture medium enhanced the cell viability. Also, effects of an amino acids (argine, aspartic acid, glutamine, glycine), as a nitrogen and carbon source in transgenic rice cell cultures were examined to enhance the cell viability and production of CTLA4Ig. Various concentrations of amino acids at 1-12 mM were added in induction phase. It was found that long-term survival of transgenic rice cells and enhanced production of hCTLA4Ig were possible by supplementing alternative energy source in transgenic rice cell cultures.