人芽囊原虫在DMEM单相培养基中体外培养方法的建立

目的建立人芽囊原虫(Blastocystis hominis,B.h)在DMEM单相培养基中的体外连续培养方法,为进一步研究其诊断、生活史、致病机制奠定基础。方法比较不同pH值、血清种类、血清浓度及接种量等条件下虫体生长繁殖情况及影响因素。结果使用DMEM培养基、接种量大于105/管、pH7.0~8.0、10%~30%小牛血清(或人、马血清),青、链霉素及二性霉素B,于37℃条件下厌氧培养,每三、六或五天转种一次,可达到体外长期培养B.h的目的。结论使用DMEM单相培养基可用于人芽囊原虫的诊断及体外长期培养。

Vitro culture of Blastocystis hominis in medium DMEM

OBJECTIVE: To establish the vitro culture of Blastocystis hominis (B. h) in medium DMEM for the further research on diagnosis, life cycle and pathogenicity of this intestinal protoza. METHODS: The growth, reproduction and relevant factors of B . h under different culture conditions including sorts and concentrations of serum, pHs and number of inoculation were compared. RESULTS: Conditions for the continuously anaerobic culture of B. h in medium DMEM were as follows: the number of inoculation were no less than 10(5) cells per tube, pHs ranged 7.0 - 8.0, concentrations of calf serum (or human serum and horse serum) ranged 10% - 30% , antibiotics and Amphotericin B should be added, subculturing could be choose at the each peaking-day 3,6 or 5 at 37 degrees C. CONCLUSION: The medium DMEM could be used in diagnosis and continuously vitro culture for B. h.