In vitro co-culture of Fasciola hepatica newly excysted juveniles (NEJs) with 3D HepG2 spheroids permits novel investigation of host–parasite interactions
Fasciola hepatica, or liver fluke, causes fasciolosis in humans and livestock. Following ingestion of vegetation contaminated with encysted parasites, metacercariae, newly excysted juveniles (NEJ) excyst in the small intestine and cross the intestinal wall. After penetrating the liver, the parasite...
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| Language: | English |
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Taylor & Francis Group
2025-12-01
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| Series: | Virulence |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/21505594.2025.2482159 |
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| author | Aiste Vitkauskaite Emma McDermott Richard Lalor Carolina De Marco Verissimo Mahshid H. Dehkordi Kerry Thompson Peter Owens Howard Oliver Fearnhead John Pius Dalton Nichola Eliza Davies Calvani |
| author_facet | Aiste Vitkauskaite Emma McDermott Richard Lalor Carolina De Marco Verissimo Mahshid H. Dehkordi Kerry Thompson Peter Owens Howard Oliver Fearnhead John Pius Dalton Nichola Eliza Davies Calvani |
| author_sort | Aiste Vitkauskaite |
| collection | DOAJ |
| description | Fasciola hepatica, or liver fluke, causes fasciolosis in humans and livestock. Following ingestion of vegetation contaminated with encysted parasites, metacercariae, newly excysted juveniles (NEJ) excyst in the small intestine and cross the intestinal wall. After penetrating the liver, the parasite begins an intra-parenchymal migratory and feeding phase that not only drives their rapid growth and development but also causes extensive haemorrhaging and immune pathology. Studies on infection are hindered by the difficulty in accessing these microscopic juvenile parasites in vivo. Thus, a simple and scalable in vitro culture system for parasite development is needed. Here, we find that two-dimensional (2D) culture systems using cell monolayers support NEJ growth to a limited extent. By contrast, co-culture of F. hepatica NEJ with HepG2-derived 3D spheroids, or “mini-livers,” that more closely mimic the physiology and microenvironment of in vivo liver tissue, promoted NEJ survival, growth, and development. NEJ grazed on the peripheral cells of the spheroids, and they released temporally regulated digestive cysteine proteases, FhCL3, and FhCL1/2, similar to in vivo parasites. The 3D co-culture induced development of the NEJ gut and body musculature, and stimulated the tegument to elaborate spines and a variety of surface sensory/tango/chemoreceptor papillae (termed S1, S2, and S3); these were especially pronounced around the oral and ventral suckers that sense host chemical cues and secure the parasite in tissue. HepG2 3D spheroid/parasite co-culture methodologies should accelerate investigations into the understanding of F. hepatica NEJ developmental biology and studies on host–parasite interactions, and streamline the search for new anti-parasite interventions. |
| format | Article |
| id | doaj-art-1da4dd5f4eeb47d2b114a8192b4fe459 |
| institution | Kabale University |
| issn | 2150-5594 2150-5608 |
| language | English |
| publishDate | 2025-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Virulence |
| spelling | doaj-art-1da4dd5f4eeb47d2b114a8192b4fe4592025-08-20T03:40:35ZengTaylor & Francis GroupVirulence2150-55942150-56082025-12-0116110.1080/21505594.2025.2482159In vitro co-culture of Fasciola hepatica newly excysted juveniles (NEJs) with 3D HepG2 spheroids permits novel investigation of host–parasite interactionsAiste Vitkauskaite0Emma McDermott1Richard Lalor2Carolina De Marco Verissimo3Mahshid H. Dehkordi4Kerry Thompson5Peter Owens6Howard Oliver Fearnhead7John Pius Dalton8Nichola Eliza Davies Calvani9Molecular Parasitology Laboratory, Centre for One Health, Ryan Institute, School of Natural Sciences, The University of Galway, Galway, The Republic of IrelandAnatomy Imaging and Microscopy (AIM), Anatomy, School of Medicine, The University of Galway, Galway, The Republic of IrelandMolecular Parasitology Laboratory, Centre for One Health, Ryan Institute, School of Natural Sciences, The University of Galway, Galway, The Republic of IrelandMolecular Parasitology Laboratory, Centre for One Health, Ryan Institute, School of Natural Sciences, The University of Galway, Galway, The Republic of IrelandPharmacology and Therapeutics, School of Medicine, The University of Galway, Galway, The Republic of IrelandAnatomy Imaging and Microscopy (AIM), Anatomy, School of Medicine, The University of Galway, Galway, The Republic of IrelandAnatomy Imaging and Microscopy (AIM), Anatomy, School of Medicine, The University of Galway, Galway, The Republic of IrelandPharmacology and Therapeutics, School of Medicine, The University of Galway, Galway, The Republic of IrelandMolecular Parasitology Laboratory, Centre for One Health, Ryan Institute, School of Natural Sciences, The University of Galway, Galway, The Republic of IrelandMolecular Parasitology Laboratory, Centre for One Health, Ryan Institute, School of Natural Sciences, The University of Galway, Galway, The Republic of IrelandFasciola hepatica, or liver fluke, causes fasciolosis in humans and livestock. Following ingestion of vegetation contaminated with encysted parasites, metacercariae, newly excysted juveniles (NEJ) excyst in the small intestine and cross the intestinal wall. After penetrating the liver, the parasite begins an intra-parenchymal migratory and feeding phase that not only drives their rapid growth and development but also causes extensive haemorrhaging and immune pathology. Studies on infection are hindered by the difficulty in accessing these microscopic juvenile parasites in vivo. Thus, a simple and scalable in vitro culture system for parasite development is needed. Here, we find that two-dimensional (2D) culture systems using cell monolayers support NEJ growth to a limited extent. By contrast, co-culture of F. hepatica NEJ with HepG2-derived 3D spheroids, or “mini-livers,” that more closely mimic the physiology and microenvironment of in vivo liver tissue, promoted NEJ survival, growth, and development. NEJ grazed on the peripheral cells of the spheroids, and they released temporally regulated digestive cysteine proteases, FhCL3, and FhCL1/2, similar to in vivo parasites. The 3D co-culture induced development of the NEJ gut and body musculature, and stimulated the tegument to elaborate spines and a variety of surface sensory/tango/chemoreceptor papillae (termed S1, S2, and S3); these were especially pronounced around the oral and ventral suckers that sense host chemical cues and secure the parasite in tissue. HepG2 3D spheroid/parasite co-culture methodologies should accelerate investigations into the understanding of F. hepatica NEJ developmental biology and studies on host–parasite interactions, and streamline the search for new anti-parasite interventions.https://www.tandfonline.com/doi/10.1080/21505594.2025.2482159Fasciola hepaticaliver flukeHepG2spheroids3D cell culturehelminth |
| spellingShingle | Aiste Vitkauskaite Emma McDermott Richard Lalor Carolina De Marco Verissimo Mahshid H. Dehkordi Kerry Thompson Peter Owens Howard Oliver Fearnhead John Pius Dalton Nichola Eliza Davies Calvani In vitro co-culture of Fasciola hepatica newly excysted juveniles (NEJs) with 3D HepG2 spheroids permits novel investigation of host–parasite interactions Virulence Fasciola hepatica liver fluke HepG2 spheroids 3D cell culture helminth |
| title | In vitro co-culture of Fasciola hepatica newly excysted juveniles (NEJs) with 3D HepG2 spheroids permits novel investigation of host–parasite interactions |
| title_full | In vitro co-culture of Fasciola hepatica newly excysted juveniles (NEJs) with 3D HepG2 spheroids permits novel investigation of host–parasite interactions |
| title_fullStr | In vitro co-culture of Fasciola hepatica newly excysted juveniles (NEJs) with 3D HepG2 spheroids permits novel investigation of host–parasite interactions |
| title_full_unstemmed | In vitro co-culture of Fasciola hepatica newly excysted juveniles (NEJs) with 3D HepG2 spheroids permits novel investigation of host–parasite interactions |
| title_short | In vitro co-culture of Fasciola hepatica newly excysted juveniles (NEJs) with 3D HepG2 spheroids permits novel investigation of host–parasite interactions |
| title_sort | in vitro co culture of fasciola hepatica newly excysted juveniles nejs with 3d hepg2 spheroids permits novel investigation of host parasite interactions |
| topic | Fasciola hepatica liver fluke HepG2 spheroids 3D cell culture helminth |
| url | https://www.tandfonline.com/doi/10.1080/21505594.2025.2482159 |
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