Derivation and characterization of matched cell lines from primary and recurrent serous ovarian cancer

Abstract

BackgroundCell line models have proven to be effective tools to investigate a variety of ovarian cancer features. Due to the limited number of cell lines, particularly of the serous subtype, the heterogeneity of the disease, and the lack of cell lines that model disease progression, there is a need to further develop cell line resources available for research. This study describes nine cell lines derived from three ovarian cancer cases that were established at initial diagnosis and at subsequent relapse after chemotherapy.MethodsThe cell lines from three women diagnosed with high-grade serous ovarian cancer (1369, 2295 and 3133) were derived from solid tumor (TOV) and ascites (OV), at specific time points at diagnosis and relapse (R). Primary treatment was a combination of paclitaxel/carboplatin (1369, 3133), or cisplatin/topotecan (2295). Second line treatment included doxorubicin, gemcitabine and topotecan. In addition to molecular characterization (p53, HER2), the cell lines were characterized based on cell growth characteristics including spheroid growth, migration potential, and anchorage independence. The in vivo tumorigenicity potential of the cell lines was measured. Response to paclitaxel and carboplatin was assessed using a clonogenic assay.ResultsAll cell lines had either a nonsense or missense TP53 mutations. The ability to form compact spheroids or aggregates was observed in six of nine cell lines. Limited ability for migration and anchorage independence was observed. The OV3133(R) cell line, formed tumors at subcutaneous sites in SCID mice. Based on IC50 values and dose response curves, there was clear evidence of acquired resistance to carboplatin for TOV2295(R) and OV2295(R2) cell lines.ConclusionThe study identified nine new high-grade serous ovarian cancer cell lines, derived before and after chemotherapy that provides a unique resource for investigating the evolution of this common histopathological subtype of ovarian cancer.

Robust high-performance nanoliter-volume single-cell multiple displacement amplification on planar substrates

“…We next performed droplet MDA on 30 single cells from the TOV2295 cell line (25), derived from a high-grade serous ovarian cancer, which is genomically unstable and has CNVs on multiple length scales. Low-depth (mean, 0.02×) WGS was performed on all 30 samples, and higher-depth (mean, 6.5×) WGS was performed on the eight samples with the highest coverage breadth calculated from the low-depth WGS data.…”

AAV9 delivering a modified human Mullerian inhibiting substance as a gene therapy in patient-derived xenografts of ovarian cancer

“…Methods that establish primary cell lines and PDXas in parallel allow for faster in vivo and in vitro testing that could direct therapeutic strategies for individual patients in real time (19,(29)(30)(31)(32), making ascites an ideal source of material for xenografts, which we distinguished from biopsy PDXs by referring to them as PDXas. Ascites can be obtained less invasively throughout the clinical course of the disease (33), are abundant and amenable to direct engraftment in a large number of mice, and often grow in tissue culture, allowing for parallel in vitro analyses. We have optimized this source of ovarian cancer cells to test the responsiveness and mechanisms of MIS sensitivity by deriving both short-term primary cell lines and PDXas, specifically from the patient population with the recurrent platinum-resistant, high-grade serous phenotype.…”

Molecular characterization of 7 new established cell lines from high grade serous ovarian cancer

“…All cell lines in our study were established from patients receiving standard treatment, providing another advantage, which other recently established or used cell lines do not have [19][20][21]. Furthermore, two of our cell lines presented respective BRCA1 and BRCA2 mutations, each leading to a truncated protein.…”

“…A considerable study on tumor heterogeneity and clonal evolution in ovarian cancer has been performed [18] using matched cell lines established at the end of the 1980s [19] and new cell line series derived from the same patient have been established and characterized [20,21]. These approaches provide new opportunities to study HGSOC.…”