Screening and optimization of cultivation factors for improving cell growth in transgenic rice cell suspension cultures

초록

Recombinant human granulocyte-macrophage colony stimulating factor (hGM-CSF) was produced and secreted from transgenic rice cell suspension cultures using a rice a-amylase (RAmy3D) expression system. In this system, recombinant hGM-CSF expression is regulated metabolically by sugar starvation. Therefore, it is possible to separate cell growth phase from production phase for the purpose of improving the recombinant hGM-CSF yield. In this study, we investigated the effects of cultivation factors, including media components and culture condition, on cell growth. In addition, main factors were screened and optimal levels was determined for cell growth using Plackett-Burman design and Box-Behnken design with Minitab software. From the point of cell density and growth rate, sucrose, nitrogen, 2,4-D and kinetin had influenced on cell growth, and each optimum concentration was 60 g/L, 35.0 mM, 0.2 mg/L, and 0.02 mg/L, respectively.