The Effect Of Medium Composions On Protoscoleces In Vitro Cultivation

Echinococcus granulosus,consisted of hydatid cyst and its contents(brood capsule, protoscoleces,daughter cyst,grand daughter and fluid of cyst,etc),grow in organs of human and other intermediate hosts in the form of cyst.The differentiation of protoscoleces correlated with genus of intermediate hosts.Protoscoleces procreated by cyst in sheep and human,and by eggs in canidae.Prepared three kinds of culture media with different components,Groupâ… :RPMI-1640 culture medium with 10%fluid of cyst,Groupâ…¡:RPMI-1640 culture medium with 10%calf serum,Groupâ…¢:RPMI-1640 culture medium;and put the same amount of newly collected protoscoIeces in to cultivate for 15 days,with the condition of 37℃and 5%CO₂.Observed the protoscoleces at the 3^th,the 9^th and the 15^th day,and results showed that the cultural duration and components of culture media affected protoscoleces growth and development. Protoscoleces of Groupâ… remained in dormancy with the highest survival ratio,and Groupâ…¡were vigorous.The Groupâ…¢displayed intermediately.Conclusion:PCS could conduce to protoscoleces growth and development,and hydatidfluid could inhibit.The 15-day cultural protoscoleces from the three group were detected by real-time PCR for the genes(eg143,egTPx and ePC1),with the non-cultural protoscoleces and Actin I gene as control groups.Results showed that the expressions of egTPx and eg143 were much higher in Groupâ…¡than in Groupâ… andâ…¢,and that of ePC1 in Groupâ… was the highest(2.1 times as much expression as in control group).Conclusion:egTPx and eg143 could conduce to protoscoleces growth and development,and ePC1 could inhibit it.The Western-blot were done for detecting the expressed products of Eg143,EgTPx and EPC1 in the 15-day protoscoleces from the three group.Results displayed:the EgTPx and Eg143 were expressed in all protoscoleces from the four groups,and the EPC1 were expressed in all protoscoleces from the three groups except Groupâ…¡.Conclusion:the EgTPx-protein and Eg143-protein could conduce to protoscoleces growth and development, and EPC1-protein could inhibit.