Subcutaneous Transplantation Assay: A Method to Transplant Melanoma Cells from a Donor to a Recipient Zebrafish Model

- During melanoma transplantation assay, cancerous cells from tumor-bearing zebrafish are transplanted into an immunocompromised recipient model to study tumor development. Begin by taking a euthanized tumor-bearing donor zebrafish and surgically resect the tumor from its body. Transfer the tumor into a Petri dish containing a suitable buffer and mince it into small pieces.

Next, triturate the tumor fragments using a pipette. This dissociates the tumor pieces into individual cells. Now, load the tumor cell suspension into a syringe. Subsequently, prep an immunocompromised recipient Casper zebrafish on a filter paper. This pigmentation-deficient, mutant variant of zebrafish lacks melanocytes and iridophores, making it transparent and versatile for imaging and transplantation experiments.

Next, inject the preloaded melanoma cell suspension subcutaneously into the region above the peritoneal cavity, midway between the hindbrain and the dorsal fin of the Casper zebrafish. Now, transfer the recipient fish into the water and monitor it to observe tumor development. The transparent body of zebrafish helps in the easy visualization of the developed tumors.

In the following protocol, we wil