Iwanaga Masashi, Adachi Yuka, Uchiyama Koudai, Tsukui Keita, Katsuma Susumu, Kawasaki Hideki
Department of Agrobiology and Bioresources, Faculty of Agriculture, Utsunomiya University, Mine-machi 350, Utsunomiya-shi, Tochigi, 321-8505, Japan,
In Vitro Cell Dev Biol Anim. 2014 Oct;50(9):792-6. doi: 10.1007/s11626-014-9781-y. Epub 2014 Jun 17.
Bombyx mori ovary-derived BmN4 cells have been successfully adapted to a commercial serum-free medium (SFM; SF900-II) by gradually reducing the serum-containing TC-100 medium content from 100 to 0% (v/v). The BmN4 cells adapted to the SFM (BmN-SFM) adhered strongly to the culture flask and showed altered cell morphology. The BmN-SFM was subcultured 200 times, and the population doubling time was 4.70 d. Infection studies showed that BmN-SFM cells were easily susceptible to B. mori nucleopolyhedrovirus (BmNPV), and both the multiplication of budded virus and the promoter activity of the polyhedrin gene in BmN-SFM cells were almost the same as those in BmN4 cells before adaptation. Additionally, mouse interleukin-3 expressed by a recombinant BmNPV was normally secreted and modified with N-linked glycans in BmN-SFM cells. These findings indicate that BmN-SFM is particularly useful for a BmNPV-based baculovirus expression vector system with serum-free conditions.
通过将含血清的TC - 100培养基含量从100%（v/v）逐渐降至0%，家蚕卵巢来源的BmN4细胞已成功适应商业无血清培养基（SFM；SF900-II）。适应SFM的BmN4细胞（BmN - SFM）牢固地附着于培养瓶，且细胞形态发生改变。BmN - SFM传代培养200次，群体倍增时间为4.70天。感染研究表明，BmN - SFM细胞对家蚕核型多角体病毒（BmNPV）易感性高，且出芽病毒的增殖以及BmN - SFM细胞中多角体蛋白基因的启动子活性与适应前的BmN4细胞几乎相同。此外，重组BmNPV表达的小鼠白细胞介素 - 3在BmN - SFM细胞中正常分泌并进行N - 糖基化修饰。这些发现表明，BmN - SFM对于基于BmNPV的无血清条件杆状病毒表达载体系统特别有用。
