Studies of Maruca vitrata cells in serum-free suspension culture

• Main Authors: Chien Yi-Ju, 簡怡如
• Other Authors: 張春生
• Format: Others
• Language: zh-TW
• Published: 102
• Online Access: http://ndltd.ncl.edu.tw/handle/51150148912597338318

碩士 === 南台科技大學 === 生物科技系 === 101 === Insect cells have been used in developing baculovirus expression system, recombinant protein, and vaccines production, in which a large number of cells for subsequent infection are needed. Therefore, this study is aimed to look for optimal Maruca vitrata insect cell cultural conditions to achieve maximum numbers cell by spinner flask. On comparison with the cell culture adding 10% serum, 1.26 folds of the maximum number of adaptation in serum-free medium was obtained. After 4-time subcultures, the maximum number of cells and specific growth rate were stabilized, leading to the result that the serum-free medium is suitable for MV cell growth. Moreover, a appropriate amount of 8% protein is helpful for cell growth. By using Taguchi method, the best nitrogen composition in serum free medium was found as glutamine 3 g L-1, soy peptone 0.5 g L-1, ploypeptone 1 g L-1, lactabumine 4 g L-1, meat peptone 1.5 g L-1, tryptone 1.5 g L-1, and protein A 6%, which could get the maximum cells number 6.43 ± 0.31×106 cells mL-1. Also using Taguchi method, the best amino acids in serum-free medium with L-cystine, L-methionine,and L-tyrosine was found both of them the optimum concentration added 0.5 g L-1, which could get the maximum cells number 5.53 ± 0.53×106 cells mL-1. Keywords: Maruca vitrata, serum-free medium suspension culture