Fluorescently Labeled T-ALL Cells Isolation: A Method to Isolate T-ALL Cells from Adult Zebrafish

- T-cell acute lymphoblastic leukemia, or T-ALL, is a type of cancer that occurs when lymphocytes fail to mature and remain as immature cells called lymphoblasts. These abnormal cells are ineffective in fighting infection. As these immature cells divide rapidly, they outnumber healthy blood cells and can travel to the organs such as the spleen, liver, and CNS.

Tumors in zebrafish are similar to those formed in humans. Therefore, zebrafish is the preferred model to generate T-ALL tumor cells.

Begin by selecting the transformed fishes that exhibit fluorescent signals, indicating the spread of tumor cells. Remove the euthanized fish's head and wash the body cavity with heparin solution to remove excess red blood cells. Now, inject a suitable buffer into the body cavity. Apply gentle pressure from outside and collect the buffer containing tumor cells in a microfuge tube.

Stain the cells with trypan blue and observe them under a microscope. Live tumor cells appear fluorescent and colorless when stained with trypan blue, as the negatively charged cell membrane excludes the negative trypan blue from entering inside the cell. Trypan blue enters dead cells through the porous membrane, stain