In vitro Cultivation and Perturbation of Cestode Parasites

Parasite was maintained by in vivo propagation of the parasite material in mice (supplied by Xiamen University Laboratory Animals Center, XMULAC). Mature and developing protoscoleces were collected from parasite material, manually picked under the microscope, and then immediately used for RNA isolation or EdU labeling. In vitro cultivation of metacestode vesicles was performed using host cell conditioned medium as previously described [26 (link)]. The growth of metacestode vesicles and the process of vesicle formation from protoscoleces were examined after 21 days and 14 days of culture, respectively as described by Cheng et al. [27 (link)].
Aurora inhibitors (MLN8237, MLN8054, MK-5108 and AZD1152-HQPA), nocodazole and hydroxyurea were supplied by Selleck Chemicals. Drugs were added into the culture medium at a final concentration as indicated. All of the drug experiments were carried out under axenic culture conditions as described before [26 (link)]. For longer periods of treatment, experiments were performed with exchange of the medium containing the same ingredients every three days.