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使用Cellpose对斑马鱼幼虫体内异种移植的人类癌细胞进行定量的方案。

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

作者信息

Köpke Karina, Lembke Carla-Sophie, Oosterhof Nynke, Dijkstra Emma S C, Paridaen Judith T M L

机构信息

European Research Institute for the Biology of Ageing, University Medical Center Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, the Netherlands.

European Research Institute for the Biology of Ageing, University Medical Center Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, the Netherlands.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103479. doi: 10.1016/j.xpro.2024.103479. Epub 2024 Dec 9.

DOI:10.1016/j.xpro.2024.103479
PMID:39656591
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11681912/
Abstract

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.

摘要

在此,我们展示了一种用于定量斑马鱼幼虫中异种移植的人胶质母细胞瘤干细胞(GSCs)的方案。我们首先描述将GSCs原位异种移植到斑马鱼幼虫中脑、免疫荧光标记和共聚焦成像的步骤。然后,我们详细介绍使用Cellpose算法进行GSC定量的程序。该方案提供了一种在异种移植实验中对人癌细胞进行半自动分割和定量的技术。通过这种方法,可以以无偏倚的方式确定癌细胞的存活和增殖情况。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/61fc5d6cd4a8/gr10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/c69bb47f6f0c/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/11c7651545c2/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/eba70d4ac6b0/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/d83138ce4be9/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/1c1ca4e9ce75/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/0dd88415cbe5/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/61a7fea0a652/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/67e0d26dde6c/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/85bbe6ec4192/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/47bc57194d41/gr9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/61fc5d6cd4a8/gr10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/c69bb47f6f0c/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/11c7651545c2/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/eba70d4ac6b0/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/d83138ce4be9/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/1c1ca4e9ce75/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/0dd88415cbe5/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/61a7fea0a652/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/67e0d26dde6c/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/85bbe6ec4192/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/47bc57194d41/gr9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/beb7/11681912/61fc5d6cd4a8/gr10.jpg

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本文引用的文献

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A quantitative characterization of early neuron generation in the developing zebrafish telencephalon.发育中斑马鱼端脑早期神经元生成的定量表征。
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Tissue clearing and 3D imaging in developmental biology.组织透明化和 3D 成像在发育生物学中的应用。
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Generation of Zebrafish Larval Xenografts and Tumor Behavior Analysis.斑马鱼幼体异种移植的构建及肿瘤行为分析
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Morphological Abnormalities and Gene Expression Changes Caused by High Incubation Temperatures in Zebrafish Xenografts with Human Cancer Cells.高温孵育条件下斑马鱼异种移植肿瘤中人源癌细胞的形态学异常和基因表达变化。
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Glioblastoma Stem Cells and Comparison of Isolation Methods.胶质母细胞瘤干细胞及分离方法比较
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