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  3. Quantification of cell death and proliferation of patient-derived ovarian cancer organoids through 3D imaging and image analysis

Quantification of cell death and proliferation of patient-derived ovarian cancer organoids through 3D imaging and image analysis

  • STAR Protoc. 2023 Nov 16;4(4):102683. doi: 10.1016/j.xpro.2023.102683.
Aikaterini Skorda 1,  Anna Røssberg Lauridsen 1,  Kaisa Huhtinen 2,  Alexandra Lahtinen 3,  Wojciech Senkowski 4,  Jaana Oikkonen 3,  Johanna Hynninen 5,  Sampsa Hautaniemi 3,  Tuula Kallunki 6
Affiliations

Affiliations

  • 1 Danish Cancer Institute, Danish Cancer Society, 2100 Copenhagen, Denmark.
  • 2 Research Program in Systems Oncology, Research Programs Unit, Faculty of Medicine, University of Helsinki, 00014 Helsinki, Finland; Institute of Biomedicine and FICAN West Cancer Centre, University of Turku and Turku University Hospital, 20014 Turku, Finland.
  • 3 Research Program in Systems Oncology, Research Programs Unit, Faculty of Medicine, University of Helsinki, 00014 Helsinki, Finland.
  • 4 Biotech Research and Innovation Centre, University of Copenhagen, 2200 Copenhagen, Denmark.
  • 5 Department of Obstetrics and Gynaecology, University of Turku and Turku University Hospital, 200521 Turku, Finland.
  • 6 Danish Cancer Institute, Danish Cancer Society, 2100 Copenhagen, Denmark; Drug Design and Pharmacology, University of Copenhagen, 2200 Copenhagen, Denmark. Electronic address: tk@cancer.dk.
PMID: 37976153 DOI: 10.1016/j.xpro.2023.102683
Abstract

Patient-derived organoids (PDOs) are ideal ex vivo model systems to study Cancer progression and drug resistance mechanisms. Here, we present a protocol for measuring drug efficacy in three-dimensional (3D) high-grade serous ovarian Cancer PDO cultures through quantification of cytotoxicity using propidium iodide incorporation in dead cells. We also provide detailed steps to analyze proliferation of PDOs using the Ki67 biomarker. We describe steps for sample processing, immunofluorescent staining, high-throughput confocal imaging, and image-based quantification for 3D cultures. For complete details on the use and execution of this protocol, please refer to Lahtinen et al. (2023).1.

Keywords

Cancer; Cell Biology; High-Throughput Screening; Organoids.

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