Effects of HNMPA(AM)
on parasite larvae. A) Primary cells were isolated from axenic metacestode vesicles and cultivated in 2% FCS/(D)MEM supplemented with 10 nM human insulin, with or without HNMPA(AM)3. After three weeks of incubation, mature vesicles were counted. Insulin was added to the cultures in order to obtain vesicle formation within three weeks. B) Formation of primary cell aggregates in the presence of HNMPA(AM)3. Primary cells were incubated as in (A) for seven days before aggregates were fixed, embedded in Technovit 8100 and 4 μm sections stained with haematoxylin/eosin. Note the profound effect of HNMPA(AM)3 on parasite aggregates already after seven days. Ctrl, DMSO control. C) Protoscoleces were treated with HNMPA(AM)3 for two weeks under axenic conditions. Protoscolex viability was analysed by counter-staining with methylene blue. D) Metacestode vesicles were treated for one week with 100 μM HNMPA(AM)3 under axenic conditions. Survival was assessed by counting physically damaged vesicles. Vesicles were incubated in the presence of conditioned medium for optimal maintenance and survival conditions. (*) P values below 0.05, (**) very significant for P between 0.001 and 0.01, (***) extremely significant for P <0.001. DMSO, dimethyl sulphoxide; HNMPA, 2-hydroxynaphthalen-1-yl-methylphosphonic acid.