Optimization and workflow of in vitro culture of adult Fasciola hepatica

The aim of this study was to evaluate different transportation and incubation conditions to test the viability of adult Fasciola hepatica in order to propose a new cultivation workflow. The adult stage of F. hepatica was obtained from naturally infected cattle at a local slaughterhouse in Lima, Peru. Different transport and incubation conditions of F. hepatica were tested, evaluating its viability through a motility scale. DMEM and RPMI 1640 media presented better transport conditions compared to Hedon-Fleig and PBS media (p < 0.001), maintaining the flukes at 37°C. Also, DMEM and RPMI-1640 media presented better incubation conditions compared to Hedon-fleig (p < 0.001). A minimum of 3 ml of medium per fluke was required to maintain best viability (p < 0.001) and no differences in viability were found between the different types of culture plates (p > 0.05). In addition, we found that incubation with DMSO (dimethyl sulfoxide) at concentrations greater than 0.5% v/v for 48 hours generates toxicity (p < 0.001). In conclusion, RPMI 1640 and DMEN media presented better transport and in vitro cultivation conditions for F. hepatica, using DMSO at concentrations lower than 0.5% v/v.