3D-Fish Cell Culture: Some Do and Some Do Not

Abstract

3-dimensional fish cell culture systems are advantageous tools to study e.g. cell interactions, cell differentiation and tissue formation. However, cultivation of 3-dimensional cultures is difficult. A lot more has to be learned about the features that allow cultivation of tissue nnnnnn-like structures, after all about the interplay of different cell types. Recently, we showed that fibroblast-like cells from rainbow trout (RT) skin obviously helped to enhance survival rates of epithelial-like cells that were suspected to be terminally differentiated. We now made the observation that the longterm RT-fibroblast-like cell cultures of the skin started to build organoid-like structures spontaneously after long-time cultivation without subcultivation. No contact inhibition was observed. Compared to other cells from RT these cells showed an enormous growth after confluence. This was documented by timelapse imaging. RT-brain-derived cells or pancreas-derived cells from sturgeon didn’t show this feature. The expression of the following markers in RTskin derived organoid bodies has been analysed by immuncytochemistry: Cytokeratin 7, 14 and 18, Collagen Type I and Pax 6. Stem cell markers Sox2 and Oct 3/4 have been analysed, too. Overall we conclude that the ability of fish cells to grow in vitro in 3D strongly depends on the cell type, the environment in vitro and the factors secreted by the cells. Fibroblast-like cells of rainbow trout apparently provide a good platform for further research on e.g. wound healing experiments and mass production of fish cells.