To begin, thaw vial containing one milliliter of frozen GFP-labeled zebrafish T-ALL cells in a 37 degree Celsius water bath with gentle shaking. Slowly pipette the contents of the vial into a 15-milliliter conical tube, containing four milliliters of fish media on ice. Spin down the cells at 2, 500 G for five minutes at four degrees Celsius and discard the supernate before resuspending the pellet in 0.5 milliliters of fish media.
Add 10 microliters of cell suspension into the microcentrifuge tube containing 10 microliters of trypan blue dye. Mix well and transfer 10 microliters onto a hemocytometer and count the cells under the microscope. Next, hold the anesthetized adult CG1 zebrafish with ventral side up.
Using a Hamilton microsyringe, inject five to six microliters of cell suspension into the intraperitoneal space. Approximately 21 to 28 days post transplant, assess the percentage of the GFP-labeled leukemia cells from anesthetized zebrafish. To harvest leukemia cells, move the euthanized zebrafish into a new Petri dish and add one milliliter of fish media to serve as a buffer for the cells.
Using a razor blade, first decapitate zebrafish, then macerate the tissue. Pipette up and down to dislodge large clumps of cells. Pass the cell suspension through a 40-micron cell strainer into a 50-milliliter conical tube to dissociate into a single cell suspension.
