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The Use of Ovarian Cancer Cells from Patients Undergoing Surgery to Generate Primary Cultures Capable of Undergoing Functional Analysis

Lookup NU author(s): Dr Rachel O'Donnell, Dr Aiste McCormick, Dr Asima Mukhopadhyay, Michelle Dixon, Dr Angelika Kaufmann, Dr San Soo Hoo, Dr Ahmed Elattar, Professor Nicola Curtin, Professor Richard Edmondson

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

The use of cell lines or animal models has significant disadvantages when dealing with a set of heterogeneous diseases such as epithelial ovarian cancer. This has clinical relevance in that biomarkers developed using cell line or animal models are often not transferable to the clinical setting. In this study, we describe the development of a robust protocol for developing primary cultures of ovarian cancer which will overcome some of these difficulties. Women undergoing surgery for ovarian cancer were recruited and samples of ascites and solid tumour deposits were used to develop primary cultures. Cells were characterised using a panel of immunofluorescent antibodies prior to use in a variety of assays including functional assessment of DNA repair pathways. During the four year study period, viable cultures, confirmed to be epithelial in origin were generated from 156 of 172 (91%) cases recruited. Characterisation was carried out using a panel of antibodies including pancytokeratin, CA125, EpCAM, MOC-31, D2-40 and vimentin. Senescence occurred between the 2nd and 8th passages in all cultures except one in which spontaneous immortalization occurred. Cells could be successfully cultured even after a period of storage at 4 degrees C and cultured cells were capable of being used for a variety of applications including functional assays. Upon functional assessment there was minimal intra-tumour heterogeneity. It is therefore possible to derive viable ovarian cancer cell cultures in the majority of patients undergoing surgery. Cells cultured directly from patient cancers provide an accurate and highly diverse model.


Publication metadata

Author(s): O'Donnell RL, McCormick A, Mukhopadhyay A, Woodhouse LC, Moat M, Grundy A, Dixon M, Kaufmann A, Soohoo S, Elattar A, Curtin NJ, Edmondson RJ

Publication type: Article

Publication status: Published

Journal: PLoS ONE

Year: 2014

Volume: 9

Issue: 3

Online publication date: 06/03/2014

Acceptance date: 02/02/2014

Date deposited: 28/05/2014

ISSN (electronic): 1932-6203

Publisher: Public Library of Science

URL: http://dx.doi.org/10.1371/journal.pone.0090604

DOI: 10.1371/journal.pone.0090604


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