Title

Comparison and Evaluation of Echinococcus Granulosus Protoscoleces Excretory/Secretory Proteins in PBS Complemented with Glucose, DMEM and RPMI Culture Media

Abstract

Background and Objective: Preparation of proper antigens is an important issue in serology of hydatidosis. Investigators have been able to obtain excretory/secretory antigens (E/S Ags) by short-term culture of protoscoleces in a couple of culture media. However, no data are available about production rate of such antigens in different culture media. The present study was carried out to evaluate the production of E/S Ags (proteins) in PBS complemented with glucose, DMEM and RPMI culture media. Material and Methods: To obtain E/S proteins, protoscoleces of echinococcus were cultured in PBS complemented with 10% glucose, RPMI and DMEM for 72 hours. Proteins secreted in culture media were concentrated and assayed. To characterize different components, proteins were electrophoresed on SDS-PAGE. Data were analyzed using One-way ANOVA and Tukey HSD tests. Results: The mean concentration of E/S proteins in PBS medium in 24 hours of culture was significantly higher than DMEM and RPMI (P<0.05). However, such a difference was not observed between E/S proteins in DMEM and RPMI media. E/S proteins obtained from PBS medium were separated into 12 major bands and the two other media into 14 major bands within a range of molecular masses of 16 to 67 kDa. Conclusion: PBS complemented with glucose is more appropriate than the two other media for E/S proteins production. The best time to obtain E/S proteins is the first 48 hours of culture.