Neves L, Cross H F, Loureiro L, Akça A, Hommel M, Trees A J
Liverpool School of Tropical Medicine, UK.
Parasitology. 2001 Oct;123(Pt 4):357-63. doi: 10.1017/s0031182001008502.
The microaerophilous stationary phase system (MASP) was introduced in 1980 and successfully used as a standard technique for Babesia bovis and B. bigemina in vitro culture. The percentage of serum in the medium and the dependence on specific serum donors have been recognized as important constraints both for immunochemical studies and for the logistics of culture routine. In the present study the supplementation of RPMI 1640 with hypoxanthine at a concentration between 50 and 200 microM has enabled patterns of growth of B. bovis and B. bigemina to be achieved comparable to the standard technique with a simultaneous reduction of serum concentration from 40% to 5%. With hypoxanthine-supplemented medium it was possible to either replace the bovine serum from a specific donor with horse serum or use commercial adult bovine serum or foetal calf serum at 10%. When the serum replacement media Albumax II and GF21 were used, the growth of both B. bovis and B. bigemina markedly decreased after 3 x 72 h cycles. However, when these species were cultivated in culture flasks previously coated with cells from a murine peritoneal lavage, continuous parasite growth was achieved.
微需氧固定相系统（MASP）于1980年被引入，并成功用作牛巴贝斯虫和双芽巴贝斯虫体外培养的标准技术。培养基中血清的百分比以及对特定血清供体的依赖性已被认为是免疫化学研究和培养常规后勤工作的重要限制因素。在本研究中，在RPMI 1640中添加浓度为50至200微摩尔的次黄嘌呤，能够实现牛巴贝斯虫和双芽巴贝斯虫的生长模式，与标准技术相当，同时血清浓度从40%降至5%。使用添加次黄嘌呤的培养基，可以用马血清替代特定供体的牛血清，或使用10%的商业成年牛血清或胎牛血清。当使用血清替代培养基Albumax II和GF21时，在3个72小时周期后，牛巴贝斯虫和双芽巴贝斯虫的生长均明显下降。然而，当这些物种在先前用小鼠腹腔灌洗细胞包被的培养瓶中培养时，实现了寄生虫的持续生长。
