BIOTEK2021-Bioteknologi for verdiskaping

Over 2 million Salmon are used annually in basal aquaculture research in Norway, constituting 87% of the research animals used. Live fish experiments are highly expensive and time-consuming and can be ethically controversial with regard to animal welfare and the environment. Whereas in vitro models employing fish cell cultures exist, these are presently considered too remote from in life conditions to replace whole-animal experiments. This is largely due to the lack of suitable scaffold materials that allow cultivation and sustained survival of primary fish cells in a three-dimensional and more physiologically relevant environment. CABARET has been a collaborative project between the partners SINTEF, SINTEF TTO, Havforskningsinstituttet and NTNU and has developed and evaluated alginate-based scaffold materials for 3D cultivation of primary cells from Atlantic salmon. Alginate forms thermally stabile hydrogels under mild conditions and is particularly well suited for cultivation of cells at low temperatures. The material and the gelling conditions can also be tailored for obtain mechanical properties optimized for a wide spectrum of cell and tissue types. Alginate can further be functionalized for biological interaction with cells. This is well established for human cells but requires continued development and optimization for fish cells. Moreover, the project has established novel and important technology to evaluate the effect of scaffold materials in small scale and for a large sample number (high-throughput screening). Using robotic systems, 3D cultures can be established, maintained and analyzed in automated protocols allowing us to screen many different scaffold materials and cultivation conditions. This has been a necessary tool to find optimal materials for fish cells and the technology is translatable to cultivation of cells and tissues from other organisms. 3D cultivation in alginate hydrogels has shown a prolonged and more stabile viability of the cells. This means that the cells collected from one fish can be used in more experiments and the 3D cultures also provide a longer time frame for analyses compared with other methods. The project has further demonstrated a higher sensitivity of cells in 3D culture toward pesticides compared with conventional 2D cultures, which is presumed to be a more physiologically accurate response. CABARET has developed important technology for more robust cell-based research models and will contribute to a future reduction of research animals used. The results from the project will be developed further for potential commercialization and will later be published in scientific and popular scientific literature.

CABARET-Prosjektet har utviklet nye materialer og metoder for 3D-kultivering av lakseceller (lever- og hodenyreceller), samt metoder for karakterisering og anvendelse av cellekulturene innen forskning på fiskehelse. Gjennom genererte vitenskapelige data og målrettet formidling har prosjektet stimulert nye samarbeid, og tilrettelagt for fremtidig samarbeid, med nasjonale forskningsmiljøer og industri. Forskningspartnerne har utviklet ny kompetanse for utvikling av mer avanserte laboratoriemodeller både innenfor fiskeceller og humane celler, som vil implementeres i pågående og nye prosjekter. I tillegg til å generere data for potensiell patentering og fremtidig kommersialisering, har prosjektet etablert nye metoder som vil publiseres i vitenskapelig tidsskrift og utvide forskningsvirksomheten til prosjektpartnerne.

The project aims to develop novel scaffolds of sulfated alginate hydrogels for 3D cultivation of primary fish cells at low temperatures; the Cabaret assay kit. The kit will sustain function and viability of cells for in vitro analyses in an in vivo-like environment. The scaffolds will be optimized for primary Atlantic salmon cells and integrated into standardized assay kits for each cell type. Proof-of-principle will be improved viability and function of cells in relevant environmental toxicity screening assays. Salmon in aquaculture research constitute 87% of research animals used in Norway. Live fish experiments are costly and time-consuming, and 2D in vitro cell culture models existing today are too remote from in vivo conditions to replace animals, as the cells need to grow in a structured microenvironment to sustain functions. Commercially available scaffold materials cannot support viability and function of salmon cells, creating a market opportunity for novel materials. Sulfated alginate hydrogels have demonstrated great promise, and the Cabaret assay kits will allow the establishment of 3D in vitro assays as cost-effective alternatives to whole-animals in fish nutrition and toxicology research. Toward this end, a library of functionalized alginates will be generated by SINTEF and NTNU, and evaluated as scaffold materials and cell culture substrates. NIFES will perform screening assays to identify the best scaffolds for each primary cell type, and the qualifying materials will be included in the Cabaret assay kits. If both 2D cell culture and live fish experiments are partially replaced by the Cabaret assay kit, it will be possible to sell 25,000 kits yearly, corresponding to annual sales of 50-100 million NOK. Patents will be secured for developed materials, technology and/or applications, and for products and services. The project will proceed over two years, at an estimated budget of 6.2 million NOK for the partners SINTEF, NIFES, NTNU and SINTEF TTO.