Vega C A, Buening G M, Rodriguez S D, Carson C A
Vet Parasitol. 1986 Dec;22(3-4):223-33. doi: 10.1016/0304-4017(86)90109-3.
The original Babesia bigemina culture conditions were modified with regard to infected bovine erythrocyte concentration and atmospheric environment. A procedure was designed which would yield a homogeneous parasite population, beginning with a single infected erythrocyte. Calculated dilutions were made in 96-well tissue culture plates to approach one infected erythrocyte per four wells. Growth of parasites in wells was detected between 16 and 28 days after cultures were initiated. Clones were transferred to 24-well tissue culture plates for regular maintenance. Three primary clones were selected for additional recloning. The probability that the parasites detected in one well are the progeny of a single infected erythrocyte approaches 0.99 for tertiary clones.
针对感染牛红细胞浓度和大气环境，对原有的双芽巴贝斯虫培养条件进行了修改。设计了一种程序，从单个感染红细胞开始，可产生均匀的寄生虫群体。在96孔组织培养板中进行计算好的稀释，使每四个孔中接近有一个感染红细胞。培养开始后16至28天检测孔中寄生虫的生长情况。将克隆转移到24孔组织培养板中进行常规维持培养。选择三个初级克隆进行进一步的再克隆。对于三级克隆，在一个孔中检测到的寄生虫是单个感染红细胞后代的概率接近0.99。
