Oellermann R A, Carter P
Onderstepoort J Vet Res. 1977 Sep;44(3):139-42.
The optimum conditions for the culture of cells from dissociated spleens were determined. Routinely, 10(7) cells were seeded per ml of RPMI 1640 medium supplemented with 20% pre-tested foetal calf serum. For the assay of the immune response, cultures were supplemented with 30 muMolar mercaptoethanol. The immune responses to sheep erythrocyte and bluetongue virus antigens were determined by the haemolytic plaque-forming cell assays described by Oellermann (1974) and Oellermann, Carter & Marx (1976a). The optimum sheep erythrocyte antigen concentration was 2 X 10(6) erythrocytes per 10(7) spleen cells and maximum IgM plaque-forming cells were detected after 4 days in culture. Successful stimulation of the immune response to bluetongue virus was achieved in spleen cell cultures from mice previously primed with bluetongue virus. The optimum antigen concentration was 30-40 ng bluetongue virus per 10(7) spleen cells and the maximum plaque-forming cell response was observed after 4 days in culture.
确定了从解离的脾脏中培养细胞的最佳条件。通常，每毫升补充有20%预先检测的胎牛血清的RPMI 1640培养基接种10⁷个细胞。为了检测免疫反应，培养物中添加了30微摩尔的巯基乙醇。通过Oellermann（1974年）和Oellermann、Carter及Marx（1976年a）描述的溶血空斑形成细胞试验来测定对绵羊红细胞和蓝舌病病毒抗原的免疫反应。最佳的绵羊红细胞抗原浓度是每10⁷个脾细胞2×10⁶个红细胞，培养4天后检测到最大的IgM空斑形成细胞。在先前用蓝舌病病毒致敏的小鼠的脾细胞培养物中成功刺激了对蓝舌病病毒的免疫反应。最佳抗原浓度是每10⁷个脾细胞30 - 40纳克蓝舌病病毒，培养4天后观察到最大的空斑形成细胞反应。
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