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. 2024 Nov 22;33(4):e014524.
doi: 10.1590/S1984-29612024064. eCollection 2024.

Optimization and workflow of in vitro culture of adult Fasciola hepatica

Cesar Burga-Cisterna  1 , Edith Málaga  2 , Enrique Serrano-Martínez  1 , Giovana Livia-Córdova  3 , Ramón Antezana  4 , Américo Castro Luna  5 , Monica Pajuelo  6
Affiliations

Optimization and workflow of in vitro culture of adult Fasciola hepatica

Cesar Burga-Cisterna et al. Rev Bras Parasitol Vet. .

Abstract
in English, Portuguese

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.

O objetivo deste estudo foi avaliar diferentes condições de transporte e incubação, para testar a viabilidade de Fasciola hepatica adulta, a fim de propor um novo fluxo de cultivo. O estágio adulto de F. hepatica foi obtido de bovinos, naturalmente infectados, em um matadouro local em Lima, Peru. Foram testadas várias condições de transporte e incubação de F. hepatica, avaliando sua viabilidade por meio de uma escala de motilidade. Os meios DMEM e RPMI 1640 apresentaram melhores condições de transporte em comparação com os meios Hedon-Fleig e PBS (p < 0,001), mantendo os vermes a 37°C. Além disso, os meios DMEM e RPMI-1640 apresentaram melhores condições de incubação em comparação ao Hedon-fleig (p < 0,001). Foi necessário um mínimo de 3 ml de meio por verme para manter a melhor viabilidade (p < 0,001) e não foram encontradas diferenças na viabilidade entre os diferentes tipos de placas de cultura (p > 0,05). Além disso, foi constatado que a incubação com DMSO (dimetilsulfóxido) em concentrações superiores a 0,5% v/v, por 48 horas, gera toxicidade (p < 0,001). Conclui-se que os meios RPMI 1640 e DMEN apresentaram melhores condições de transporte e cultivo in vitro para F. hepatica, utilizando DMSO em concentrações inferiores a 0,5% v/v.

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Conflict of interest statement

Conflict of interest: The authors declare that they have no conflict of interest.

Figures

Figure 1
Figure 1. In vitro culture workflow of adult stage of Fasciola hepatica.
See this image and copyright information in PMC

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