Zebrafish Xenograft Model for Pancreatic Cancer

For zebrafish xenografts, wild type Tübingen (TU) zebrafish were bred and maintained in the Western Australian Zebrafish Experimental Research Centre (Biomedical Research Facility- Shenton Park, Western Australia). Experiments and data analyses were done as previously described [32 (link)]. Briefly, HPAF-II human pancreatic cancer cells were incubated with Vibrant™-Dil dye (ThermoFisher Scientific) 4 μL/mL in HBSS at 37 °C for 10 min, followed by 15 min on ice in the dark. Cells were then harvested and resuspended at a density of 10⁷ cells/, loaded into a capillary glass needle using a puller (p-97 Flaming/Brown by Sutter Instrument®) and 10 nL of cell suspension (approximately 100 cells/embryo) was injected in the perivitelline space of 24-h post fertilisation (hpf) zebrafish embryos. Zebrafish were incubated at 34 °C O/N to allow for cell growth and the following day embryos were equally distributed in to three treatment groups. One group did not receive any treatment or cells (blank), a second group was treated with 0.1% DMSO and a third group was treated with 10 μM JVG045. At 5 days post fertilisation (dpf), the effect of the drugs on cancer cell growth was documented using a fluorescent stereomicroscope equipped with a digital camera (Nikon SMZ Zoom). Images were analysed using ImageJ. Non injected embryos were used to subtract background fluorescence.