{"title":"Organoid-Immune Cell Co-culture for Stable Live Imaging.","authors":"Nathalia Ferreira, Frauke Alves, Andrea Markus","doi":"10.1007/7651_2025_627","DOIUrl":null,"url":null,"abstract":"<p><p>Patient-derived organoids (PDOs) have emerged as a promising model for personalized drug testing. Generated from human tumor samples, PDOs effectively recapitulate the genetic and phenotypic heterogeneity of patient tumors, making them an ideal ex vivo platform for studying therapeutic responses, particularly to chemotherapies. However, their lack of components of the immune system limits their use in immunotherapy testing. The following protocol facilitates the co-culture of PDOs from tumor tissue with HLA-matched peripheral blood mononuclear cells (PBMCs) in a fixed Z-plane for stable live-cell imaging. This three-dimensional co-culture method represents a significant advancement in enabling real-time assessment of immunotherapeutic effects on tumor-derived PDOs by live cell imaging.</p>","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Methods in molecular biology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1007/7651_2025_627","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 0
Abstract
Patient-derived organoids (PDOs) have emerged as a promising model for personalized drug testing. Generated from human tumor samples, PDOs effectively recapitulate the genetic and phenotypic heterogeneity of patient tumors, making them an ideal ex vivo platform for studying therapeutic responses, particularly to chemotherapies. However, their lack of components of the immune system limits their use in immunotherapy testing. The following protocol facilitates the co-culture of PDOs from tumor tissue with HLA-matched peripheral blood mononuclear cells (PBMCs) in a fixed Z-plane for stable live-cell imaging. This three-dimensional co-culture method represents a significant advancement in enabling real-time assessment of immunotherapeutic effects on tumor-derived PDOs by live cell imaging.
期刊介绍：
For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.
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