Davis W C, Talmadge J E, Parish S M, Johnson M I, Vibber S D
Infect Immun. 1978 Nov;22(2):597-602. doi: 10.1128/iai.22.2.597-602.1978.
Bovine erythrocytes infected with Anaplasma marginale were cultured for 1 to 5 days in a CO2 incubation chamber, pulse-labeled with [3H]thymidine and [14C]methionine, lysed, and fractionated by differential centrifugation and continuous density gradient centrifugation in Renografin. Anaplasma and associated fragments of stroma formed two distinct bands in the dense region of the gradient. Electron microscopic examination of pelleted material from the bands from cells cultured for 1 day revealed the presence of organisms that were morphologically intact or in various states of degeneration. Examination of fractions from the gradient for incorporation of label revealed that analplasma present in erythrocytes can incorporate both [3H]thymidine and [14C]methionine. Subsequent experiments demonstrated that organisms cultured for 3 and 5 days incorporated the radiolabeled compounds also, but to a lesser extent. The experiments demonstrate that it is possible to culture analplasma in vitro for short periods of time and monitor their growth characteristics.
将感染了边缘无形体的牛红细胞在二氧化碳培养箱中培养1至5天，用[3H]胸腺嘧啶核苷和[14C]甲硫氨酸进行脉冲标记，裂解后，通过差速离心和在泛影葡胺中进行连续密度梯度离心进行分级分离。无形体和相关的基质碎片在梯度的高密度区域形成两条明显的带。对培养1天的细胞的带中的沉淀物质进行电子显微镜检查，发现存在形态完整或处于各种退化状态的生物体。对梯度分级分离物进行标记掺入检查发现，红细胞中存在的无形体可以掺入[3H]胸腺嘧啶核苷和[14C]甲硫氨酸。随后的实验表明，培养3天和5天的生物体也能掺入放射性标记化合物，但程度较低。这些实验表明，在体外短时间培养无形体并监测其生长特性是可能的。
