McIntosh A H, Christian P D, Grasela J J
USDA, Agricultural Research Service, Biological Control of Insects Research Laboratory, Columbia, Missouri, USA.
In Vitro Cell Dev Biol Anim. 1999 Feb;35(2):94-7. doi: 10.1007/s11626-999-0007-7.
A total of eight cell lines were established from Helicoverpa armigera (3) and H. punctigera (5) embryos and ovaries. Cell lines were established and grown in TC100 and/or TC199-MK containing 10% fetal bovine serum. The serum-free medlium ExCell 400 was also used, with and without 10% supplemental fetal bovine serum, but failed to generate cell lines from fat bodies, embryos, or ovarian tissues. Cell lines consisted of heterogenous cell types ranging from oval to fibroblast-like. This is the first report on the successful establishment of cell lines from H. punctigera. Cell lines from the two species were distinguishable from each other by DAF-PCR, and noticeable differences in minor bands were observed among cell lines from the same species. All of the established cell lines from both species were susceptible to HzSNPV but did not replicate more virus than that of a H. zea cell line (BCIRL-HZ-AM1-A11). However, an H. punctigera cell line (HP1) replicated AcMNPV to the highest titer (1.0 X 10(8) 50% tissue culture infective dose/ml), and only one of the H. armigerm cell lines (HA1) was susceptible to this virus.
总共从棉铃虫(3个)和烟芽夜蛾(5个)的胚胎及卵巢中建立了8个细胞系。细胞系在含有10%胎牛血清的TC100和/或TC199-MK中建立并培养。还使用了无血清培养基ExCell 400,添加和不添加10%补充胎牛血清,但未能从脂肪体、胚胎或卵巢组织中生成细胞系。细胞系由从椭圆形到成纤维细胞样的异质细胞类型组成。这是关于成功从烟芽夜蛾建立细胞系的首次报道。通过DAF-PCR可区分这两个物种的细胞系,并且在同一物种的细胞系之间观察到小条带存在明显差异。从这两个物种建立的所有细胞系都对HzSNPV敏感,但与玉米螟细胞系(BCIRL-HZ-AM1-A11)相比,复制的病毒并不更多。然而,一个烟芽夜蛾细胞系(HP1)将AcMNPV复制到最高滴度(1.0×10⁸ 50%组织培养感染剂量/毫升),而棉铃虫细胞系中只有一个(HA1)对这种病毒敏感。