A simple, quick and economic method for in vitro cultivation of Echinococcus multilocularis metacestode and generation of primary cells

Abstract

Abstract Background Alveolar echinococcosis is considered to be one of the most potentially lethal parasitic zoonotic diseases. However, the molecular mechanisms by which Echinococcus multilocularis interact with host are poorly understood and only limited treatments available. Due to the great advantages of cell culture systems for molecular research, numerous attempts have been made to establish primary cell cultures for Echinococcus multilocularis. In this study, we developed a simple, rapid and economic method, which allows Echinococcus multilocularis metacestodes tissue blocks to generate daughter vesicles without the presence of host feeder cells in regular medium. Methods We performed anaerobic, hypoxic (1% O2), normoxic and semi-anaerobic (in sealed tubes) cultures for Echinococcus multilocularis metacestodes tissues to produce daughter vesicles. Then the daughter vesicles were cultured at three distinct oxygen consentrations (anaerobic, 1% O2, normoxic) to search optimal cultivation conditons. The daughter vesicle’s viability was assayed by carboxyfluorescein succinimidyl ester staining. These vesicles with good vigor were subsequently used for testing antiparasitic effect of albendazole, isolating primary cells and infecting animals. Results After 4 weeks incubation, we found that Echinococcus multilocularis metacestodes tissues only cultured in sealed tubes produced daughter vesicles. And the daughter vesicles were observed remarkably enlarged under anaerobic conditions after 8 days of culture, while vesicles cultured under other two conditions showed a mild increase in volume. Our in vitro cultivated vesicles had good viability and can be used for testing of antiparasitic drugs, isolating primary cells and infecting animals. Conclusions In the present work we established a simple, quick and economic method for in vitro generation vesicles from tissue blocks of Echinococcus multilocularis metacestodes in the absence of host feeder cells. Our in vitro cultivated vesicles with good viability are suitable for screening drugs for treatment of alveolar echinococcosis in vitro and in vivo.