title: Primary cell culture of Echinococcus granulosus developed from the cystic germinal layer: biological and functional characterization.
abstract: Cell cultures of parasitic helminths are an invaluable tool for investigations of basic biological processes, as well as for development of improved chemotherapeutic agents and molecular interactions between host and parasite. We carried out a simple and efficient methodology to isolate Echinococcus granulosus germinal cells which were maintained for at least 4 months while cultivated in the presence of reducing agents and hormones. Microscopic analysis of the primary cell culture revealed the presence of cells with similar Echinococcus germinal cell morphology and behaviour. Population doubling time was estimated at 48 h, showing a rapid division rate. To discard possible host contamination, the specificity of the primary culture was tested by nested PCR, analyzing mdh gene expression and obtaining only one product with the expected size. We also studied the expression of specific E. granulosus proteins in primary cell culture. The novel and systematized method described here constitutes a powerful tool for investigations in cystic echinococcosis on biochemical and biological aspects related to the life cycle of the parasite and mechanisms of host-parasite interactions. This method also constitutes a powerful tool for the design of more efficient therapeutic alternatives.
authors.affiliation: Laboratorio de Zoonosis Parasitarias, Departamento de Biología, Facultad de Ciencias Exactas y Naturales, Universidad Nacional de Mar del Plata, Funes 3350, Nivel Cero, 7600 Mar del Plata, Argentina. calbani@mdp.edu.ar
authors.forename: Clara M
authors.initials: CM
authors.lastname: Albani
authors.forename: María Celina
authors.initials: MC
authors.lastname: Elissondo
authors.forename: Andrea C
authors.initials: AC
authors.lastname: Cumino
authors.forename: Andrea
authors.initials: A
authors.lastname: Chisari
authors.forename: Guillermo M
authors.initials: GM
authors.lastname: Denegri
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000469:parasitology*; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D003560:Cysts / Q000469:parasitology*; D004443:Echinococcosis / Q000469:parasitology / Q000662:veterinary*; D048209:Echinococcus granulosus / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification / Q000378:metabolism; D015801:Helminth Proteins / Q000235:genetics / Q000378:metabolism; D008018:Life Cycle Stages; D008099:Liver / Q000469:parasitology; D008168:Lung / Q000469:parasitology
chemical_list: D015801:Helminth Proteins