Abstract A cell line derived from Spodoptera litura, SL7B, was used to ascertain mass production of Autograph californica nucleopolyhedrovirus (AcMNPV). Results revealed that the SL7B cell line was better than a commercial cell line, SF21AE, for the replication of wild type AcMNPV and for the amount of recombinant virus protein expressed. SL7B cells can grow in low-cost and simple ISC-03 medium, and growth rates increased upon an increase in concentration of fetal bovine serum (FBS) contained in the culture medium. In addition, the production of wild type AcMNPV and amounts of the recombinant protein, luciferase, expressed had a positive correlation to the concentration of FBS in the medium. Addition of Pluronic F68 in ISC-03 was helpful to the growth of SL7B cells. When Pluronic F-68 was added, the cell growth rate was faster in medium containing 2% FBS than in medium containing 8% FBS. Moreover, in medium containing the same concentration of FBS, both the wild type AcMNPV and the recombinant protein production rate were much higher when supplemented with Pluronic F-68. In addition, the multiplicity of infection (MOI) and timing of infection were also very important for using the cell culture to produce virus. The greatest yield of virus production was with an MOI = 0.1 at the initiation (day 0) stage of cells cultured in TNM-FH medium. However, the best cell growth in ISC-03 medium was obtained with an MOI = 10 at 3 d after culturing.
摘要 本文利用斜紋夜蛾細胞株SL7B進行病毒量產試驗，發現就細胞株種類而言，斜紋夜蛾細胞株比現行商品化秋行軍蟲細胞株SF21AE更適合用來生產苜蓿夜蛾核多角體病毒。利用簡易培養液ISC-03培養SL7B細胞時，隨培養液中添加血清濃度增加，細胞生長速度加快且細胞生產野生型病毒數量與表現重組病毒外源蛋白產量均增加。Pluronic F-68添加物對細胞生長有益，在含2%胎牛血清的ISC-03培養液中添加0.1% 的Pluronic F-68，則細胞生長速度比添加8%胎牛血清者更快；當培養液中血清濃度相同時，Pluronic F-68的添加不但可增加細胞生產病毒核多角體數量且可提高重組蛋白螢光酵素表現量。此外，細胞培養生產病毒時，病毒感染劑量(multiplicity of infection, MOI)與感染時機相當重要，以TNM-FH + 8% FBS培養時，在第0天以MOI = 0.1感染，可得最高病毒產量；而以ISC-03 + 8% FBS培養時，則在第3天以高MOI = 10感染所得產量最高。
| Key words: Spodoptera litura cell line, AcMNPV, recombinant protein expression, cell population doubling time, medium supplements. | | 關鍵詞: 斜紋夜蛾細胞株，苜蓿夜蛾核多角體病毒，重組蛋白產量，細胞倍增時間，細胞培養液添加物 |
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