Optimization of yeast-derived serum-free medium by Plackett-Burman design and response surface methodology
10.13200/j.cnki.cjb.003906
- VernacularTitle:Plackett-Burman实验和响应面法优化酵母源无血清培养基
- Author:
XIAO Meng
- Publication Type:Journal Article
- Keywords:
Yeast extract(YE);
CHO cell;
Serum-free medium(SFM);
Plackett-Burman design;
Response surface methodology(RSM);
Optimization
- From:
Chinese Journal of Biologicals
2024;37(8):989-995+1001
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo optimize the nutrient components of yeast-derived serum-free medium(SFM)and determine the composition and concentration by Plackett-Burman design and response surface methodology(RSM).MethodsBased on the SFM selected in the early stage of experiment,10 nutrient components were screened by PBD,using Chinese hamster ovary(CHO)cells as the research object,which were optimized secondarily by RSM to elucidate the interaction among the variables and determine the best medium composition for the growth of CHO cells,after the factors promoting the growth of CHO cells were screened out. CHO cells in logarithmic growth stage were re-suspended in the optimized culture medium,and then subcultured and amplified.ResultsAmong the 10 variables screened by Plackett-Burman experiment,yeast extract(YE)FM888 and trace elements effectively increased the specific growth rate of cells;Non-essential amino acids and FM888 showed significant effects on improving cell survival days;FM888,essential amino acids,non-essential amino acids and trace elements significantly increased the density of viable cells. Based on this,three factors,FM888,trace elements and non-essential amino acids,were comprehensively selected for RSM optimization test. When cultured in the optimized medium in suspension,the maximum specific growth rate of CHO cells was 0. 025/h,the survival days were 6d,and the maximum viable cell density was 7. 187 × 10~6cells/mL. The optimized medium was used for cell passage test,based on which,the cells were stably passaged for 12 passages,and the viable cell density and cell viability were close to those of the control group(commercial SFM).ConclusionThe yeast-derived SFM with clear components and supporting the high-density growth of CHO cells was obtained,which provided a reference and basis for the application of YE in animal cell culture and the development of high-efficiency SFM.
