Optimization and workflow of in vitro culture of adult Fasciola hepatica

Burga-Cisterna C.; Málaga E.; Serrano-Martínez E.; Livia-Córdova G.; Antezana R.; Luna A.C.; Pajuelo M. · Revista Brasileira de Parasitologia Veterinaria (2024) · DOI: 10.1590/s1984-29612024064

Resumen

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.

Instituciones: Universidad Peruana Cayetano Heredia; Universidad Nacional Mayor de San Marcos; Universidad Nacional Pedro Ruiz Gallo