Zebrafish Tumor Graft Transplantation to Grow Tumors In Vivo That Engraft Poorly as Single Cell Suspensions

Abstract

Angiosarcoma is a clinically aggressive tumor with a high rate of mortality. It can arise in vascular or lymphatic tissues, involve any part of the body, and aggressively spread locally or metastasize. Angiosarcomas spontaneously develop in the tp53 deleted (tp53^del/del) zebrafish mutant. However, established protocols for tumor dissection and transplantation of single cell suspensions of angiosarcoma tumors result in inferior implantation rates. To resolve these complications, we developed a new tumor grafting technique for engraftment of angiosarcoma and similar tumors in zebrafish, which maintains the tumor microenvironment and has superior rates of engraftment.

Keywords: angiosarcoma; tp53; tumor graft; zebrafish; zebrafish cancer model.

FIG. 1.

Engraftment and growth of angiosarcoma graft in syngeneic CG1 zebrafish. (A) Growth of GFP+ angiosarcoma tumor over time (scale bar = 3 mm). (B) Representative H&E staining of the engrafted angiosarcoma tumor (40 × , scale bar = 60 μm). (C) Graph shows the exponential growth of the normalized angiosarcoma tumor volume over time of the fish in (A). (D) Comparison between number of zebrafish with engrafted tumors, as well as time (days) to engraftment, between the two transplantation methods (single cell suspensions versus tumor grafts) for both angiosarcoma and MPNSTs. GFP, green fluorescent protein; MPNSTs, malignant peripheral nerve sheath tumors. H&E, hematoxylin and eosin.