猪附红细胞体的体外培养研究

Research on the in Vitro Maintenance of Mycoplasma suis(Eperythrozoon suis)

用RPMI-1640和胎牛血清按比例混合并添加肌苷作为体外培养猪附红细胞体(M.suis)的基础培养基,以无附红细胞体感染的兔红细胞泥作为培养寄生载体,置于体积分数为5%CO2的37℃生化恒温培养箱进行M.suis的体外培养研究.结果表明,培养时每24 h换培养液1次,每36 h传代1次,可连续传代40代次以上,并能保持最高感染率90%以上;感染M.suis的猪红细胞在4℃条件下可保持30 d以上而不发生溶血;兔红细胞接种新鲜制备的自然感染M.suis的猪红细胞,36 h后感染率可达91.6%,并一直维持到96 h,96 h后有少量细胞开始出现溶血;接种4℃条件下保存30 d以内自然感染M.suis的猪红细胞与新鲜制备的自然感染M.suis的猪红细胞对兔红细胞的感染率影响差异不显著.

In vitro culture of Mycoplasma suis （Eperythrozoon suis ） was carried out by using the medium prepared with the ratios of RPMI-1640 supplemented with 40% Fetal Bovine Serum （FBS） ,0.2% inosine and rabbit red blood cell （RBC） acting as the in vitro culture carrier to multiplicate the Mycoplasma suis（ M. suis）. The in vitro maintenance of M. suis was proceeded in the 37 ℃ constant temperature and 5% CO2 condition. The result showed that the highest infection rates of rabbit RBC could be up to above 90% and the in vitro incubation could be successively passed over 40 times by changing the medium every 24 h and passing one time every 36 h. At 36 h post-inoculation of the newly prepared porcine RBC naturally infected M. suis, the peak infection ratio of about 91.6% was reached and maintained until 96 h post-inoculation. The germfree naturally infected M. suis porcine RBC could be stored at 4 % for 30 d without hemolysis. All the M. suis grew well before 96 h in the incubation process, and only a few rabbit RBC hemolysis was happened after 96 h. No obvious different rabbit RBC infection ratio was displayed by comparing with inoculating naturally infected M. suis porcine RBC with the newly prepared and stored at 4 ℃ before 30 d.