摘要
本研究对目前应用最多的两种昆虫细胞BTI-Tn-5B1-4和Sf-9在无血清培养基Sf-900Ⅲ中进行驯化和培养,现已稳定传至45代以上。比较了两种细胞在Sf-900Ⅲ和有血清培养基TNM-FH中的细胞形态、生长速率、病毒产量和重组蛋白表达水平。结果表明,在Sf-900Ⅲ中Sf-9细胞比TNM-FH培养的略大,BTI-Tn-5B1-4的圆形细胞增多,BTI-Tn-5B1-4和Sf-9在Sf-900Ⅲ中生长速度快,群体倍增时间分别为18.5h和21.7h,而在TNM-FH中,BTI-Tn5B1-4和Sf-9的群体倍增时间分别为21.9h和25.4h;在两种培养基中BTI-Tn-5B1-4和Sf-9细胞的病毒感染率、多角体产量和β-半乳糖苷酶(β-galactosidase)的表达水平均无明显差异,而碱性磷酸酶(secreted alkaline phosphatase,SEAP)在Sf-900Ⅲ中的表达水平明显高于TNM-FH中的水平。
BTI -Tn -5B1 -4 and Sf- 9 cell lines, that are dominant cell lines with basic study and application in the world, were adapted to a serum -free medium, Sf- 900 m for approximately 45 passages. The characteristics including morphology, growth kinetics, virus production and recombinant protein expression were compared with that of they grew in Sf- 900 m and TNM -FH. The results showed that the two cells line in Sf- 900Ⅲ were little bigger than in TNM -FH. The cell population doubling times of both BTI -TnSB1 -4 and Sf- 9 cells in Sf- 900Ⅲ(18.5 and 21.7 h, respectively) shortened in comprision to that in TNM -FH(21.9 h for BTI- TnSB1 -4 and 25.4 h for Sf - 9). The virus infection rate, production of polyhedra and expression of β - galactosidase (β - gal) in both types of media were not significantly different, but the expression of secreted alkaline phosphatase (SEAP) in Sf- 900Ⅲ medium was higher than in serum -containing cultures.
出处
《青岛农业大学学报(自然科学版)》
2009年第2期131-135,共5页
Journal of Qingdao Agricultural University(Natural Science)
基金
国家自然科学基金(30771451)
国家973计划(2009CB118902)
关键词
昆虫细胞
无血清培养
生长曲线
病毒产量
蛋白表达
cell lines
serum - free culture
growth kinetics
virus production
protein expression