Vaughn J L, Fan F
Insect Biocontrol Laboratory, USDA, Beltsville, Maryland 20705, USA.
In Vitro Cell Dev Biol Anim. 1997 Jun;33(6):479-82. doi: 10.1007/s11626-997-0067-5.
The development of a serum-free medium that supports the growth of cells from a Spodoptera frugiperda and a Lymantria dispar cell line is reported. A yeast hydrolysate provided the B-vitamin complex, and a combination of a meat hydrolysate and tryptose provided most of the free amino acids required for cell growth. Supplemental cystine and methionine were required to achieve maximum cell growth. The serum or serum replacements used in earlier formulations were replaced with commercial lipid preparations and increased levels of iron salts. Although the cell growth cycle had a somewhat extended lag phase and the population doubling time of the S. frugiperda cells was longer than on serum-containing medium, the saturation densities were much higher. Spodoptera cells grown in this medium replicated the Autographa californica nuclear polyhedrosis virus well, producing 8.71 x 10(6) TCID50 extracellular virus and 4.4 x 10(6) polyhedra/ml culture. The specific activity of the polyhedra was somewhat less than that of polyhedra produced in insects.
报道了一种无血清培养基的研发，该培养基可支持来自草地贪夜蛾和舞毒蛾细胞系的细胞生长。酵母水解物提供了B族维生素复合物，肉水解物和胰蛋白胨的组合提供了细胞生长所需的大部分游离氨基酸。需要补充胱氨酸和蛋氨酸以实现细胞的最大生长。早期配方中使用的血清或血清替代物被商业脂质制剂和增加的铁盐水平所取代。尽管细胞生长周期的延迟期有所延长，且草地贪夜蛾细胞的群体倍增时间比在含血清培养基上更长，但饱和密度要高得多。在此培养基中生长的草地贪夜蛾细胞能很好地复制苜蓿银纹夜蛾核型多角体病毒，产生8.71×10(6) TCID50的细胞外病毒和4.4×10(6)个多角体/毫升培养物。多角体的比活性略低于在昆虫体内产生的多角体。
