Understanding the ways in which pathogens infect host cells is essential to improve and develop new treatment strategies. This study aimed to generate a novel in vitro infection model by establishing a reproducible 3D spheroid cell culture system that may lead to a reduced need for animals in fish disease research. 2D models (commonly cell lines) cannot replicate many key conditions of in vivo infections, but 3D spheroids have the potential to provide bridging technology between in vivo and in vitro systems. 3D spheroids were generated using cells from rainbow trout (Oncorhynchus mykiss) cell lines, RTG-2 and RTS-11. The RTG-2 spheroids were tested for their potential to be infected upon exposure to Saprolegnia parasitica spores. Positive infiltration of mycelia into the spheroids was verified by confocal microscopy. As a closer analogue of in vivo conditions encountered during infection, the straightforward model developed in this study shows promise as an additional tool that can be used to further our understanding of host–pathogen interactions for Saprolegnia and possibly a variety of other fish pathogens.
Bibliographical noteFunding Information:
JMS was supported by an Erasmus Mundus Scholarship. MF was supported by a PhD studentship funded from the EU H2020 (H2020‐SFS‐10a‐2014) programme (ParaFishControl, 634429). MS, CJS and PvW were supported by the BBSRC (BB/M026566/1 & BB/P020224/1). Thanks also go to Dr Dawn Shewring and Kevin Mackenzie for their excellent technical assistance.
- 3D cell culture
- fish model
- Parasite culture