{"title":"Protocol for formation, staining, and imaging of 3D breast cancer models using MicroTissues mold systems.","authors":"Ben Haspels, Maayke M P Kuijten","doi":"10.1016/j.xpro.2024.103250","DOIUrl":null,"url":null,"abstract":"<p><p>The use of 3D cancer models to study therapy response has a great advantage over conventional 2D cell culture. Here, we present an optimized protocol for breast cancer spheroid and rod-shaped microtissue formation using MicroTissues mold systems. We describe steps for cast formation and treating the 3D models with DNA-damaging agents. We then detail procedures for analyzing the 3D models by whole-mount immunostaining and confocal imaging of fixed samples or with the use of live-cell reporters.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"5 4","pages":"103250"},"PeriodicalIF":1.3000,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11459068/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"STAR Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.xpro.2024.103250","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/9/20 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
Abstract
The use of 3D cancer models to study therapy response has a great advantage over conventional 2D cell culture. Here, we present an optimized protocol for breast cancer spheroid and rod-shaped microtissue formation using MicroTissues mold systems. We describe steps for cast formation and treating the 3D models with DNA-damaging agents. We then detail procedures for analyzing the 3D models by whole-mount immunostaining and confocal imaging of fixed samples or with the use of live-cell reporters.
与传统的二维细胞培养相比，使用三维癌症模型研究治疗反应具有很大的优势。在此，我们介绍了使用 MicroTissues 模具系统形成乳腺癌球形和杆形微组织的优化方案。我们介绍了铸模形成和用 DNA 破坏剂处理三维模型的步骤。然后，我们详细介绍了通过整装免疫染色和共聚焦成像固定样本或使用活细胞报告来分析三维模型的步骤。
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