Perfusion-based ex vivo culture of frozen ovarian cancer tissues with preserved tumor microenvironment

Abstract Ovarian cancer (OC) poses significant treatment challenges due to late-stage diagnosis and a complex tumor microenvironment contributing to therapy resistance. We optimized a U-CUP perfusion-based bioreactor method to culture patient-derived primary and metastatic OC specimens, demonstratin...

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Main Authors: Monica De Luise, Ivana Kurelac, Sara Coluccelli, Antonio De Leo, Ewelina M. Bartoszek, Maria Iorio, Marco Grillini, Camelia Alexandra Coadă, Dario de Biase, Lorena Marchio, Mónica Núñez López, Natalie Rimmer, Anna Myriam Perrone, Pierandrea De Iaco, Anna Maria Porcelli, Viola Heinzelmann, Ivan Martin, Francis Jacob, Manuele Giuseppe Muraro, Giuseppe Gasparre
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:npj Precision Oncology
Online Access:https://doi.org/10.1038/s41698-025-00941-6
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author Monica De Luise
Ivana Kurelac
Sara Coluccelli
Antonio De Leo
Ewelina M. Bartoszek
Maria Iorio
Marco Grillini
Camelia Alexandra Coadă
Dario de Biase
Lorena Marchio
Mónica Núñez López
Natalie Rimmer
Anna Myriam Perrone
Pierandrea De Iaco
Anna Maria Porcelli
Viola Heinzelmann
Ivan Martin
Francis Jacob
Manuele Giuseppe Muraro
Giuseppe Gasparre
author_facet Monica De Luise
Ivana Kurelac
Sara Coluccelli
Antonio De Leo
Ewelina M. Bartoszek
Maria Iorio
Marco Grillini
Camelia Alexandra Coadă
Dario de Biase
Lorena Marchio
Mónica Núñez López
Natalie Rimmer
Anna Myriam Perrone
Pierandrea De Iaco
Anna Maria Porcelli
Viola Heinzelmann
Ivan Martin
Francis Jacob
Manuele Giuseppe Muraro
Giuseppe Gasparre
author_sort Monica De Luise
collection DOAJ
description Abstract Ovarian cancer (OC) poses significant treatment challenges due to late-stage diagnosis and a complex tumor microenvironment contributing to therapy resistance. We optimized a U-CUP perfusion-based bioreactor method to culture patient-derived primary and metastatic OC specimens, demonstrating that perfusion better preserves cancer cell viability and proliferation, both when fresh and slow-frozen tissues were used. Perfused cultures maintained key microenvironment components, including cancer-associated fibroblasts, endothelial and immune cells. Genetic analysis confirmed the retention in culture of tumor-specific driver mutations. We hence challenged ad hoc generated cisplatin-sensitive and resistant OC cells with cisplatin during growth in U-CUP, validating our system for the testing of drug response. Finally, treatment of slow-frozen OC tissues with carboplatin/paclitaxel revealed different degrees of response to treatment, as indicated by variations in tumor necrosis and number of residual PAX8+ cells, providing the bases for the prompt evaluation of OC standard chemotherapy efficacy in our ex vivo system.
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spelling doaj-art-dcd2efcb9c114a4b9b934ba65ac96d0a2025-08-20T01:52:24ZengNature Portfolionpj Precision Oncology2397-768X2025-05-019111410.1038/s41698-025-00941-6Perfusion-based ex vivo culture of frozen ovarian cancer tissues with preserved tumor microenvironmentMonica De Luise0Ivana Kurelac1Sara Coluccelli2Antonio De Leo3Ewelina M. Bartoszek4Maria Iorio5Marco Grillini6Camelia Alexandra Coadă7Dario de Biase8Lorena Marchio9Mónica Núñez López10Natalie Rimmer11Anna Myriam Perrone12Pierandrea De Iaco13Anna Maria Porcelli14Viola Heinzelmann15Ivan Martin16Francis Jacob17Manuele Giuseppe Muraro18Giuseppe Gasparre19Department of Medical and Surgical Sciences (DIMEC), University of BolognaDepartment of Medical and Surgical Sciences (DIMEC), University of BolognaSolid Tumor Molecular Pathology Laboratory, IRCCS Azienda Ospedaliero-Universitaria di BolognaDepartment of Medical and Surgical Sciences (DIMEC), University of BolognaMicroscopy Core Facility, Department of Biomedicine, University of BaselDepartment of Medical and Surgical Sciences (DIMEC), University of BolognaPathology Unit, IRCCS Azienda Ospedaliero-Universitaria di BolognaDepartment of Medical and Surgical Sciences (DIMEC), University of BolognaSolid Tumor Molecular Pathology Laboratory, IRCCS Azienda Ospedaliero-Universitaria di BolognaDepartment of Medical and Surgical Sciences (DIMEC), University of BolognaOvarian Cancer Research, Department of Biomedicine, University of Basel and University Hospital of BaselOvarian Cancer Research, Department of Biomedicine, University of Basel and University Hospital of BaselDepartment of Medical and Surgical Sciences (DIMEC), University of BolognaDepartment of Medical and Surgical Sciences (DIMEC), University of BolognaIRCCS Azienda Ospedaliero-Universitaria di BolognaOvarian Cancer Research, Department of Biomedicine, University of Basel and University Hospital of BaselTissue Engineering, Department of Biomedicine, University of Basel and University Hospital of BaselOvarian Cancer Research, Department of Biomedicine, University of Basel and University Hospital of BaselTissue Engineering, Department of Biomedicine, University of Basel and University Hospital of BaselDepartment of Medical and Surgical Sciences (DIMEC), University of BolognaAbstract Ovarian cancer (OC) poses significant treatment challenges due to late-stage diagnosis and a complex tumor microenvironment contributing to therapy resistance. We optimized a U-CUP perfusion-based bioreactor method to culture patient-derived primary and metastatic OC specimens, demonstrating that perfusion better preserves cancer cell viability and proliferation, both when fresh and slow-frozen tissues were used. Perfused cultures maintained key microenvironment components, including cancer-associated fibroblasts, endothelial and immune cells. Genetic analysis confirmed the retention in culture of tumor-specific driver mutations. We hence challenged ad hoc generated cisplatin-sensitive and resistant OC cells with cisplatin during growth in U-CUP, validating our system for the testing of drug response. Finally, treatment of slow-frozen OC tissues with carboplatin/paclitaxel revealed different degrees of response to treatment, as indicated by variations in tumor necrosis and number of residual PAX8+ cells, providing the bases for the prompt evaluation of OC standard chemotherapy efficacy in our ex vivo system.https://doi.org/10.1038/s41698-025-00941-6
spellingShingle Monica De Luise
Ivana Kurelac
Sara Coluccelli
Antonio De Leo
Ewelina M. Bartoszek
Maria Iorio
Marco Grillini
Camelia Alexandra Coadă
Dario de Biase
Lorena Marchio
Mónica Núñez López
Natalie Rimmer
Anna Myriam Perrone
Pierandrea De Iaco
Anna Maria Porcelli
Viola Heinzelmann
Ivan Martin
Francis Jacob
Manuele Giuseppe Muraro
Giuseppe Gasparre
Perfusion-based ex vivo culture of frozen ovarian cancer tissues with preserved tumor microenvironment
npj Precision Oncology
title Perfusion-based ex vivo culture of frozen ovarian cancer tissues with preserved tumor microenvironment
title_full Perfusion-based ex vivo culture of frozen ovarian cancer tissues with preserved tumor microenvironment
title_fullStr Perfusion-based ex vivo culture of frozen ovarian cancer tissues with preserved tumor microenvironment
title_full_unstemmed Perfusion-based ex vivo culture of frozen ovarian cancer tissues with preserved tumor microenvironment
title_short Perfusion-based ex vivo culture of frozen ovarian cancer tissues with preserved tumor microenvironment
title_sort perfusion based ex vivo culture of frozen ovarian cancer tissues with preserved tumor microenvironment
url https://doi.org/10.1038/s41698-025-00941-6
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