20.500.12556/RUL-146346 Refined high-content imaging-based phenotypic drug screening in zebrafish xenografts Zebrafish xenotransplantation models are increasingly applied for phenotypic drug screening to identify small compounds for precision oncology. Larval zebrafish xenografts offer the opportunity to perform drug screens at high-throughput in a complex in vivo environment. However, the full potential of the larval zebrafish xenograft model has not yet been realized and several steps of the drug screening workflow still await automation to increase throughput. Here, we present a robust workflow for drug screening in zebrafish xenografts using high-content imaging. We established embedding methods for high-content imaging of xenografts in 96-well format over consecutive days. In addition, we provide strategies for automated imaging and analysis of zebrafish xenografts including automated tumor cell detection and tumor size analysis over time. We also compared commonly used injection sites and cell labeling dyes and show specific site requirements for tumor cells from different entities. We demonstrate that our setup allows us to investigate proliferation and response to small compounds in several zebrafish xenografts ranging from pediatric sarcomas and neuroblastoma to glioblastoma and leukemia. This fast and cost-efficient assay enables the quantification of anti-tumor efficacy of small compounds in large cohorts of a vertebrate model system in vivo. Our assay may aid in prioritizing compounds or compound combinations for further preclinical and clinical investigations. cancer models phenotypic screening fenotipski pregled cebrica tumorske celice presajanje zdravil ksenografti celic onkologija farmacevtska kemija tumorji true false true Angleški jezik Slovenski jezik Članek v reviji 2023-05-24 10:41:11 2023-05-24 10:41:17 2025-08-08 13:22:22 0000-00-00 00:00:00 2023 0 0 16 str. art. 44 Vol. 7 2023 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 615.4:54:616-006 2397-768X 10.1038/s41698-023-00386-9 153170179 s41698-023-00386-9.pdf s41698-023-00386-9.pdf 1 D5259D55451AA25C4EBB20F6EF2066EB 599e8c94ebc47907d72f9556736f1feb4382cc73c66e326b2926431370693de6 13a67a3f-fa11-11ed-a314-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=169509 https://www.nature.com/articles/s41698-023-00386-9 1 7904f5be-fa0e-11ed-a314-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=169508 Fakulteta za farmacijo 0 0 0