Velichkova M, Simeonov I, Petkov P G, Iotov M
Vet Med Nauki. 1984;21(6):28-33.
Attempts were made to produce multilayer cell cultures of chick fibroblasts in a RC-42 roller as well as to replicate in these the Aujezsky's disease virus. Used were fibroblasts of 10-11-day-old chick embryos. The cells were obtained in a closed system after the classic method with an electromagnetic stirrer and a 0.3 percent trypsin in phosphate buffer, having no Ca and Mg ions. Hank's solution was used as a nutrient growth medium, containing 0.5 per cent lactoalbumin hydrolysate and calf serum at pH 7.4-7.6. Both the cell growth and the cytopathic effect of the virus in the stationary and the roller cultures were carried out simultaneously through microscopy, and the evaluation was made after a fourgrade system. It was found that proper cell cultures or chick fibroblasts could be obtained under the following optimal parameters: volume of nutrient medium--200 cm.3; amount of normal inactivated calf sera--10 per cent; and speed of rotation--10 r. p. m. The titer of the virus was found to surpass by 2 logarithms the virus in the stationary cell cultures for a twice shorter period of cultivation.
尝试在RC - 42转瓶中制备鸡成纤维细胞多层培养物,并在其中复制奥捷斯基氏病病毒。使用的是10 - 11日龄鸡胚的成纤维细胞。细胞是按照经典方法,在封闭系统中,用电磁搅拌器和不含钙镁离子的磷酸盐缓冲液中的0.3%胰蛋白酶获得的。汉克氏溶液用作营养生长培养基,含有0.5%的水解乳白蛋白和小牛血清,pH值为7.4 - 7.6。通过显微镜同时观察病毒在静置培养和转瓶培养中的细胞生长及细胞病变效应,并按照四级系统进行评估。结果发现,在以下最佳参数下可获得合适的细胞培养物或鸡成纤维细胞:营养培养基体积——200立方厘米;正常灭活小牛血清量——10%;转速——每分钟10转。发现在培养时间缩短一半的情况下,病毒滴度比静置细胞培养中的病毒滴度高出2个对数。