U251HF-GFP cells, grown to 70–80% confluence were transplanted (25–50 cells/animal) in the vicinity of the midbrain-hindbrain boundary of 36 hours post-fertilization tricaine-anesthetized zebrafish embryos (Sigma-Aldrich, St Louis, MO) and larvae screened at 24 h post-transplantation to determine implantation (27 (link)). Five days after transplantation (5 dpt) animals (24 animals/group) were randomized to vehicle (DMSO), onalespib at 0.5 μM, temozolomide (TMZ) at 10 μM or a combination of onalespib and TMZ in fish water for 5 days (from 5–10 dpt). After 5 days of treatment, tumor burden was assessed by imaging live fish on an Andor spinning disc confocal microscope as described previously (27 (link)).
Protocol detail
Zebrafish Xenograft Model for Drug Testing
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After treatment
Animals
Cells
Confocal microscope
Dmso
Embryos
Fertilization
Fish
Hindbrain
Implantation
Larvae
Midbrain
Onalespib
Temozolomide
Transplantation
Tricaine
Tumor burden
Zebrafish
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Variable analysis
independent variables
- Treatment with vehicle (DMSO)
- Treatment with onalespib at 0.5 μM
- Treatment with temozolomide (TMZ) at 10 μM
- Treatment with a combination of onalespib and TMZ
- Tumor burden assessed by imaging live fish on an Andor spinning disc confocal microscope
- U251HF-GFP cells transplanted (25–50 cells/animal) in the vicinity of the midbrain-hindbrain boundary of 36 hours post-fertilization tricaine-anesthetized zebrafish embryos
- Screening of animals at 24 h post-transplantation to determine implantation
- Treatment duration of 5 days (from 5–10 dpt)
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