Protocol for quantifying xenografted human cancer cells in zebrafish larvae using Cellpose

作者：Karina Köpke, Carla‐Sophie Lembke, Nynke Oosterhof, Emma S C Dijkstra, Judith T.M.L. Paridaen · 发表于：STAR Protocols · 年份：2024 · DOI：10.1016/j.xpro.2024.103479 · 被引用次数：2 · 研究领域：Cell Image Analysis Techniques、Zebrafish Biomedical Research Applications、Single-cell and spatial transcriptomics

Here, we present a protocol for quantifying xenografted human glioblastoma stem cells (GSCs) in zebrafish larvae. We first describe steps for orthotopic xenotransplantation of GSCs into the midbrain of zebrafish larvae, immunofluorescent labeling, and confocal imaging. We then detail procedures for GSC quantification using the Cellpose algorithm. This protocol provides a technique for the semi-automatic segmentation and quantification of human cancer cells in xenograft experiments. With this approach, cancer cell survival and proliferation can be determined in an unbiased manner. • Orthotopic xenograft of human glioblastoma cells into live zebrafish larvae • Steps for whole-mount immunostaining and confocal imaging of zebrafish larvae • Instructions for quantification of xenografted cells using the Cellpose algorithm Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Here, we present a protocol for quantifying xenografted human glioblastoma stem cells (GSCs) in zebrafish larvae. We first describe steps for orthotopic xenotransplantation of GSCs into the midbrain of zebrafish larvae, immunofluorescent labeling, and confocal imaging. We then detail procedures for GSC quantification using the Cellpose algorithm. This protocol provides a technique for the semi-automatic segmentation and quantification of human cancer cells in xenograft experiments. With this approach, cancer cell survival ...