title: The use of ovarian cancer cells from patients undergoing surgery to generate primary cultures capable of undergoing functional analysis.
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°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.
authors.affiliation: Northern Gynaecological Oncology Centre, Queen Elizabeth Hospital, Gateshead, United Kingdom; Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Rachel L
authors.initials: RL
authors.lastname: O Donnell
authors.affiliation: Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Aiste
authors.initials: A
authors.lastname: McCormick
authors.affiliation: Northern Gynaecological Oncology Centre, Queen Elizabeth Hospital, Gateshead, United Kingdom; Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Asima
authors.initials: A
authors.lastname: Mukhopadhyay
authors.affiliation: Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Laura C
authors.initials: LC
authors.lastname: Woodhouse
authors.affiliation: Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Madeleine
authors.initials: M
authors.lastname: Moat
authors.affiliation: Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Anna
authors.initials: A
authors.lastname: Grundy
authors.affiliation: Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Michelle
authors.initials: M
authors.lastname: Dixon
authors.affiliation: Northern Gynaecological Oncology Centre, Queen Elizabeth Hospital, Gateshead, United Kingdom; Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Angelika
authors.initials: A
authors.lastname: Kaufman
authors.affiliation: Northern Gynaecological Oncology Centre, Queen Elizabeth Hospital, Gateshead, United Kingdom; Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: San
authors.initials: S
authors.lastname: Soohoo
authors.affiliation: Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Ahmed
authors.initials: A
authors.lastname: Elattar
authors.affiliation: Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Nicola J
authors.initials: NJ
authors.lastname: Curtin
authors.affiliation: Faculty Institute for Cancer Sciences, University of Manchester, Manchester, United Kingdom.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Edmondson
mesh_terms: D000951:Antigens, Neoplasm / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology; D000077216:Carcinoma, Ovarian Epithelial; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D004260:DNA Repair; D004354:Drug Screening Assays, Antitumor; D005260:Female; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D009375:Neoplasms, Glandular and Epithelial / Q000378:metabolism / Q000473:pathology* / Q000601:surgery; D010051:Ovarian Neoplasms / Q000378:metabolism / Q000473:pathology* / Q000601:surgery; D010641:Phenotype; D061251:Primary Cell Culture; D013048:Specimen Handling; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents; D007211:Indoles; C531549:rucaparib