Establishment Of A New Method For In Vitro Culture Of Echinococcus Multilocularis Larva

Alveolar echinococcosis is a zoonosis caused by larval forms of Echinococcus multilocularis,and caused serious impact on human and animal health.At present,there is a lack of methods for the in vitro culture of Echinococcus multilocularis protoscoleces.In this study,we used special medium,which added serum,hepatocyte growth factor,epidermal growth factor,insulin,amino acids,vitamins,to develop a new method for protoscoleces in vitro culture.In addition,the expression of genes involve in regulate the de-differentiation of protoscoleces was analyzed.The results showed that based on the special culture medium,a method for protoscoleces in vitro culturing was successfully established.In the special experimental culture medium group,the protoscoleces could de-differentiation into vesicles with the diameter of 3.7-8.66 on the 42 th day.However,the protoscoleces cultured in conventional medium could develop into the macroscopic vesicles with the diameter of 0.8-1.8 mm on the 42 th day,and a large number of vesicles ruptured after cultured for 42 days.In addition,the vesicles are infectious.After inoculated the vesicles into the abdomina of the mice,they can develop into Echinococcus multilocularis metacestode(larval)tissue within a large number of protoscoleces.In addition,expression of ALA2.1,KADL.2,MGST2.1,galactokinase,AB11 C.10,Exostosin 1,Syntaxin gene up-regulated in vesicles compared to protoscoleces.The expression of PARP6.3 and MGST2.1 was higher in the protoscoleces than in the vesicles.This research would offer references for control and treatment of the Echinococcosis,and also provide a new method to study the host-parasites interactions.