Waghela S D, Cruz D, Droleskey R E, DeLoach J R, Wagner G G
Department of Veterinary Pathobiology, Texas A&M University, College Station 77843, USA.
Vet Parasitol. 1997 Dec 15;73(1-2):43-52. doi: 10.1016/s0304-4017(97)00045-9.
Primary cultures of Anaplasma marginale infected erythrocytes were used to determine conditions for in vitro cultivation of the rickettsia. The infected erythrocytes that were maintained by regular addition of Glasgow's MEM with fetal calf serum and uninfected erythrocytes showed a 1-5% increase in percent infected erythrocytes on the evaluation of Giemsa stained smears. This increase in parasitemia resulted in up to 70% change in the number of infected erythrocytes. Co-culture of the infected erythrocytes with endothelial cell monolayers allowed for longer maintenance with the parasitemia ranging from 5-13% through four passages over 16 weeks. Examination of cultures using transmission electron microscopy showed initial bodies within the erythrocytes at 10 days after the initial passage of the primary culture. The endothelial cell monolayers in the co-cultures contained multiple initial bodies. We have demonstrated that A. marginale can be grown for a limited number of passages in the co-culture system, which will facilitate the development of a continuous culture of the organism.
利用边缘无形体感染的红细胞原代培养物来确定立克次体体外培养的条件。通过定期添加含胎牛血清的格拉斯哥改良伊格尔培养基维持的感染红细胞和未感染红细胞，在吉姆萨染色涂片评估中显示感染红细胞百分比增加了1% - 5%。这种寄生虫血症的增加导致感染红细胞数量变化高达70%。将感染红细胞与内皮细胞单层共培养，可维持更长时间，在16周内传代4次，寄生虫血症范围为5% - 13%。使用透射电子显微镜检查培养物发现，原代培养首次传代10天后红细胞内有初始小体。共培养中的内皮细胞单层含有多个初始小体。我们已经证明，边缘无形体可以在共培养系统中传代有限次数，这将有助于该生物体连续培养的发展。
