Abstract 3697: Optimization of human cancer cell xenografts into zebrafish larvae for high-throughput drug screening

Abstract

The use of zebrafish in cancer xenograft models has grown rapidly in recent years. This is primarily due to the fact that this model takes advantage of the ease of in vivo imaging and the high-throughput screening capabilities that zebrafish have to offer. However, researchers have yet to come to a consensus on standardized procedure to utilize zebrafish for xenograft of human cells. This study aims to optimize a zebrafish xenografting protocol for various human cancers with the intention of performing high-throughput drug screening. We tested the survival of 3 different zebrafish strains when incubated at 28°C, 34°C and 37°C. There was no difference in survival between the AB, SAT, and Casper strains of zebrafish at any temperature. Human leukemia, breast, lung, colon, and brain cancer cell lines were fluorescently labelled with Vybrant DiI cell staining dye and injected at a concentration of 100, 250, 500, or 1000 cells per 2nL droplet volume into dechorionated 2-day-old zebrafish embryos. The injection sites used were the brain, caudal vein, duct of Cuvier (DC), eye, pericardium, perivitelline space (PVS), and yolk sac. As expected, injections became more difficult and time consuming when using higher cell counts as the injector needle clogged more often. Similarly, injection sites that required more precision like the caudal vein or eye took the longest to inject successfully and had lower engraftment rates than sites like the yolk or DC. We determined that using 500 cells per injection was optimal as engraftment level was comparable to 1,000 cell injections but had improved survival and reduced injection time. Optimal injection site varied between cell lines, but overall, injections into the duct of Cuvier resulted in consistently high survival and engraftment rates with one of the lowest injection times. We are in the process of determining the extent to which the injection site affects chemotherapy response on human cells implanted into zebrafish. We are also preforming a high-throughput drug screen on human leukemia cells implanted into zebrafish to provide proof-of-principle that these methods are useful in identifying novel anti-cancer compounds. In total, this work will establish standard operating procedures for use of xenograft in zebrafish, providing new opportunities in personalized medicine and drug discovery. Citation Format: Stephen R. Dockins, Jessica S. Blackburn, Meghan G. Haney, Bradley Wilson. Optimization of human cancer cell xenografts into zebrafish larvae for high-throughput drug screening [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3697.