人芽囊原虫体外纯培养法的改良及形态观察

Modification of axenic culture medium and morphological observation of Blastocystis hominis in vitro

以琼脂取代蛋白对传统的洛克氏液-鸡蛋-血清(Locke’s egg serum,LES)双相培养基进行改良,联合采用LES双相培养基、改良LES双相培养基(m LES双相培养基)、IMDM液相培养基和IMDM固相培养基建立人芽囊原虫的体外纯培养的标准流程,并观察虫体在不同培养基中的形态特征和生长情况。结果显示,人芽囊原虫在4种培养基中均以空泡型和颗粒型的形态为主,培养后镜下未见细菌生长。在mLES双相培养基中的生长速度最快,培养后第2天原虫数为3.09×10~6,达到中对数期。原虫在LES双相培养基、mLES双相培养基和IMDM液相培养基中的生长对数期的分裂时间分别为(49.72±1.35)、(24.16±2.53)和(36.3±1.5)h。提示联合采用LES双相培养基、mLES双相培养基、IMDM液相培养基和IMDM固相培养基可建立人芽囊原虫的无菌纯培养体系。

The Locke＇s egg serum （LES） biphasic medium was modified by agar replacement of protein. A sterile in vitro culture system for Blastocystis hominis was then established using LES, modified LES （mLES） and liquid-state and solid-state Iscove＇s Modified Dulbecco＇s Medium （IMDM）. Morphological and growth characteristics of the parasites in different media were observed. Blastocystis hominis showed vacuolar and granular morphologies in all types of media and had the most rapid growth in the mLES, in which the log phase was achieved on day 2 of culture （3.09 × 10^6 parasites）. The doubling times were （49.72 ± 1.35）h in LES, （24.16 ± 2.53）h in mLES, and （36.30 ± 1.50）h in IMDM. The sterile in vitro culture system for Blastocystis hominis is established using LES, mLES, and liquidus and solid-state IMDM.