Geburtshilfe Frauenheilkd 2022; 82(10): e148
DOI: 10.1055/s-0042-1757013
Abstracts | DGGG

Isolation and characterization of extracellular vesicles (EVs) from ovarian cancer ascites – a feasibility study

Authors

  • N Reinhardt

    1   University Medical Center Schleswig Holstein, Department of Gynecology and Obstetrics, Kiel, Deutschland

  • M Lettau

    2   University Medical Center Schleswig Holstein, Institute of Immunology, Kiel, Deutschland

  • G Hugendieck

    1   University Medical Center Schleswig Holstein, Department of Gynecology and Obstetrics, Kiel, Deutschland

  • S Neumann

    1   University Medical Center Schleswig Holstein, Department of Gynecology and Obstetrics, Kiel, Deutschland

  • C Rogmans

    1   University Medical Center Schleswig Holstein, Department of Gynecology and Obstetrics, Kiel, Deutschland

  • JP Weimer

    1   University Medical Center Schleswig Holstein, Department of Gynecology and Obstetrics, Kiel, Deutschland

  • I Flörkemeier

    1   University Medical Center Schleswig Holstein, Department of Gynecology and Obstetrics, Kiel, Deutschland

  • N Arnold

    1   University Medical Center Schleswig Holstein, Department of Gynecology and Obstetrics, Kiel, Deutschland

  • P Arnold

    3   Friedrich-Alexander University, Institute of Functional and Clinical Anatomy, Erlangen, Deutschland

  • O Janssen

    2   University Medical Center Schleswig Holstein, Institute of Immunology, Kiel, Deutschland

  • N Maass

    1   University Medical Center Schleswig Holstein, Department of Gynecology and Obstetrics, Kiel, Deutschland

  • DO Bauerschlag

    1   University Medical Center Schleswig Holstein, Department of Gynecology and Obstetrics, Kiel, Deutschland

  • N Hedemann

    1   University Medical Center Schleswig Holstein, Department of Gynecology and Obstetrics, Kiel, Deutschland
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Objective Chemotherapy resistance is a major factor causing high mortality rates in ovarian cancer (OvCa). Our group recently showed that proteases play a role in mediating OvCa resistance to platinum-based chemotherapy. OvCa is often accompanied by ascites formation representing the major route of metastases. Ascites establishes a complex microenvironment including extracellular vesicles (EVs) mediating intercellular communication. Notably, OvCa-derived EVs are also released at high levels upon platinum-based chemotherapy.

The aim of this study was to establish a protocol for EV isolation from ascites to search for proteases and other potential biomarkers in ascites-derived EVs of OvCa patients.

Material and Methods A protocol for the efficient enrichment of EVs from ascites fluid was established using differential centrifugation and ultracentrifugation procedures. Particles were characterized based on Minimal Information for Studies of Extracellular Vesicles (MISEV) 2018 guidelines by Nanoparticle Tracking Analysis, Western blotting and Transmission Electron Microscopy. Protease levels were quantified by Western blotting, ELISA and flow cytometry.

Results Particles with a size of approximately 100 – 400 nm were reliably isolated. These particles were identified as EVs according to the MISEV criteria. Moreover, proteases were detected in EV lysates and on the EV surface. Protease concentrations correlated with EV count and EV marker expression (CD63).

Summary Isolation of EVs from ascites fluid is feasible. EVs of malignant patient-derived ascites contain proteases and might thus contribute to chemotherapy resistance in ovarian cancer.


Interessenkonflikt

Ich erkläre als korrespondierender Autor, dass meine Koautoren mir mitgeteilt haben, dass sie während der letzten 3 Jahre keine wirtschaftlichen oder persönlichen Verbindungen im oben genannten Sinne hatten. Auch ich selbst hatte keine derartigen Verbindungen in den letzten 3 Jahren.

Publication History

Article published online:
11 October 2022

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