Chapman W G, Ramshaw I A
Appl Microbiol. 1971 Jul;22(1):1-5. doi: 10.1128/am.22.1.1-5.1971.
The adaptation of the pig kidney cell line IB-RS-2, clone 60, to growth in suspension culture is described. When fully adapted, an approximate threefold increase in viable cells was obtained within 72 hr from initial cell concentrations of 5 x 10(5) per ml in culture volumes up to 1,500 ml. The monolayer cells (99th passage level) used to initiate the suspension cultures and the fully adapted suspension cells were shown to have an aneuploid chromosome karyotype, whereas earlier monolayer cultures (32nd passage level) had a pseudodiploid karyotype. Replicate virus titrations in monolayers prepared from suspension-adapted cells, IB-RS-2 monolayer cells, BHK monolayer cells, and in suckling mice showed that the suspension cells had retained sensitivity to foot-and-mouth disease virus. The geometric mean peak infectivity of seven strains of foot-and-mouth disease virus grown in IB-RS-2 suspension cells was 10(8.2) plaque-forming units per ml, with a mean complement-fixing activity of approximately 135 complement-fixing units per ml. These preliminary results indicate that submerged cultures of these cells on an industrial scale may be useful for commercial foot-and-mouth disease vaccine production.
本文描述了猪肾细胞系IB-RS-2克隆60适应悬浮培养生长的过程。完全适应后，在高达1500毫升的培养体积中，从每毫升5×10⁵个初始细胞浓度开始，72小时内活细胞数量增加了约三倍。用于启动悬浮培养的单层细胞（第99代水平）和完全适应的悬浮细胞显示具有非整倍体染色体核型，而早期的单层培养物（第32代水平）具有假二倍体核型。对由适应悬浮培养的细胞、IB-RS-2单层细胞、BHK单层细胞制备的单层以及乳鼠进行的重复病毒滴定表明，悬浮细胞对口蹄疫病毒仍保持敏感性。在IB-RS-2悬浮细胞中培养的七种口蹄疫病毒株的几何平均峰值感染性为每毫升10⁸·²个空斑形成单位，平均补体结合活性约为每毫升135个补体结合单位。这些初步结果表明，在工业规模上对这些细胞进行深层培养可能有助于商业化生产口蹄疫疫苗。
