Optimization of in vitro models to study the development of Echinococcus granulosus at the celular and molecular levels

PRADA L; SPILLIOTIS M; CUCHER M; BREHM K; ROSENZVIT M

Congreso; XXIII International Congress of Hydatidology; 2009

Optimization of in vitro models to study the development of Echinococcus granulosus at the cellular and molecular levels Prada L1, Spiliotis M3, Cucher M, 1Brehm K2, Rosenzvit M1 1Departamento de Microbiología, Facultad de Medicina, UBA, Buenos Aires, Argentina. 5411-59599500 ext.2192. 2Universidad de Würzburg, Alemania.3Universidad de Berna, Suiza lprada@fmed.uba.ar The study of helminth parasites has not reached the development achieved in unicellular parasites, mainly because of the lack of appropriate in vitro models. One of our goals was to optimize E. granulosus in vitro models to obtain protoscoleces (PEs) differentiated towards the strobilar adult stage or vesicular stage (microcyst). PEs were extracted from porcine hydatid cysts (pig strain) and co-cultured with different cell lines. When co-cultured with RH- cells (rat hepatocytes) PEs differentiated towards the strobilar adult stage, the appearance of banding and the subsequent formation of a third proglottid in some individuals was observed. On the other hand, PEs co-cultured with COS-7 cells vesicularized and some of them formed laminar layer. The expression of genes related with development in those stages is being analysed by RT-PCR. Additionally, we succeeded in establishing a primary cell culture of E. granulosus from the disintegration of the germinal layer of porcine hydatid cysts, which in contact with RH- cells proliferated and formed aggregates. These aggregates assembled into structures with movement and morphology similar to immature protoscoleces unlike E. multilocularis primary cells which were shown to form metacestode vesicles. The parasitic origin of the cells was determined by PCR and RT-PCR with E. granulosus specific primers. These in vitro models pave the way for the study of molecular and cellular processes in the development of E. granulosus.