Establishment Model Of Cultural In Vitro For Echinococcus And Observation Affinity With Cells

Objective: To establish model of cultural in vitro for Echinococcus with Bel7404 cell,Caco-2 cell , Hela cell and their super- natant fluid, and observation affinity with the cells. Methods: After recovery Bel-7404 cells,Caco-2 cell, Hela cell, and reserve its super- natant fluid,metacestode tissue placed in DMEM medium containing the cells and their supernatant fluid respectively was cultured in an incubator containing 5% CO2 at 37℃for a period of 38 days. Follow-up parameters were recorded during cultivation included the counting, growth, measure- ment diameters, and drawing growth curve of secondary vesicles. After the culture , the secondary vesicles'hydatid fluid and wall were detected with protein level and DNA identification. Secondary vesicles'hydatid fluid protein band were observated by SDS-PAGE.Results: Echino- coccus multilocularis metacestodes could develop in DMEM medium containing Bel-7404 cells,Caco-2 cell , Hela cell, and its superna- tant fluid in vitro. Budding of new cysts was observed and the vesicles'diameter was from 0.5 to 5mm. Detection of secondary vesicles'hydatid fluid protein level in vitro were Bel-7404 cells ,3.8 mg/ml;Bel-7404 cells'supernatant fluid,1.1 mg/ml;Caco-2 cell,3.2 mg/ml;Caco-2 cells'supernatant fluid, 1.2 mg/ml;Hela cell,2.5 mg/ml;Hela cells'supernatant fluid,3.8 mg/ml . PCR amplification of vesicles'wall DNA showed the same specific 200bp band as Echinococcus multilocu- laris.It was conformed that cultural in vitro and in vivo secondary vesicles'hydatid fluid have similar 14,47-66kd protein band by SDS-PAGE. Conclusion: Firstly, Echinococcus multilocularis metacestodes growth in vitro did not rely on co- cultural cells, completely. Secondly, different culture environment and cells in vitro resulted in different protein level. Thirdly, Echinococcus multilocularis metacestodes'affinity with Bel-7404 cells and was different from another cells.The fourth, cultural in vitro and in vivo secondary vesicles'hydatid fluid have similar proteins. Last, establishment model of cultural in vitro for Echinococcus with Bel7404 cell,Caco-2 cell , Hela cell and their super- natant fluid was feasible,however, it has some variability .