A rapid and simple method for the purification of tumor cells from ascitic fluid of ovarian carcinoma

Hirte, Hal W.; Da, Clark; Mazurka, John; O'Connell, G.; Rusthoven, James J.

doi:10.1016/0090-8258(92)90046-l

Cited by 21 publications

(10 citation statements)

References 2 publications

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“…Sometimes it was possible to obtain enough cells for analysis by filtering ascites cells through a 30-pm nylon mesh (Spectrum), which retains tumor clumps but not the smaller leukocytes, fibroblasts, or mesothelial cells. Filters were backwashed to yield essentially only tumor cell clumps (29,31). Results with this technique were similar to those obtained by fractionating cells by differential attachment.…”

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confidence: 50%

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Telomerase activity in human ovarian carcinoma.

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Hirte

Bacchetti

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

594

292

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Telomeres fulfill the dual function of protecting eukaryotic chromosomes from illegitimate recombination and degradation and may aid In chromosome attachment to the nuclear membrane. We have previously shown that telomerase, the enzyme which synthesizes telomeric DNA, is not detected in normal somatic cells and that telomeres shorten with replicative age. In cells immortalized in vitro, activation of telomerase apparently stabilizes telomere length, preventing a critical destabilization of chromosomes, and cell proliferation continues even when telomeres are short. In vivo, telomeres of most tumors are shorter than telomeres of control tissues, suggesting an analogous role for the enzyme. To assess the relevance of telomerase and telomere stability in the development and progression of tumors, we have measured enzyme activity and telomere length in metastatic cells of epithelial ovarian carcinoma. We report that extremely short telomeres are maintaine in these cells and that tumor cells, but not isogenic nonmalignant cells, express telomerase. Our findilgs suggest that progression of malignancy is ultimately dependent upon activation of telomerase and that telomerase inhibitors may be effective antitumor drugs.Telomeres contain both DNA and protein that together appear to stabilize the ends ofeukaryotic DNA (reviewed in refs.

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“…Ascitic fluid was withdrawn from ovarian carcinoma patients at the time of diagnostic laparotomy or by subsequent paracentesis (28) and centrifuged to obtain a cell pellet as described (29). Leukocytes were obtained from the pooled buffy coats ofthree normal males, and normal ovarian epithelium was obtained from the surface of the ovary (30).…”

mentioning

confidence: 99%

Telomerase activity in human ovarian carcinoma.

Counter

Hirte

Bacchetti

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

594

292

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“…Several methods for the culture of primary ovarian cancer cells isolated from ascites have been described [8]. These methods however require complex multi-step procedures.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2014

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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.

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“…Isolation and primary culturing of tumor cells from ascites have become more widespread and several different methods have been established to achieve this aim (5–8). A widely used protocol for the propagation of EOC tumor cells was developed by Langdon et al (6, 9).…”

Section: Development Of Primary Models Of Hgs Diseasementioning

confidence: 99%

Modeling Platinum Sensitive and Resistant High-Grade Serous Ovarian Cancer: Development and Applications of Experimental Systems

Cunnea

Stronach

2014

Front. Oncol.

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High-grade serous ovarian cancer remains the most common sub-type of ovarian cancer and, characterized by high degrees of genomic instability and heterogeneity, is typified by a transition from early response to acquired resistance to platinum-based chemotherapy. Conventional models for the study of ovarian cancer have been largely limited to a set of relatively poorly characterized immortalized cell lines and recent studies have called into question the validity of some of these as reliable models. Here, we review new approaches and models systems that take into account advances in our understanding of ovarian cancer biology and advances in the technology available for their generation and study. We discuss primary cell models, 2D, 3D, and organotypic models, and “paired” sample approaches that capture the evolution of chemotherapy failure within single cases. We also overview new methods for non-invasive collection of representative tumor material from blood samples. Adoption of such methods and models will improve the quality and clinical relevance of ovarian cancer research.

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A rapid and simple method for the purification of tumor cells from ascitic fluid of ovarian carcinoma

Cited by 21 publications

References 2 publications

Telomerase activity in human ovarian carcinoma.

Telomerase activity in human ovarian carcinoma.

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

Modeling Platinum Sensitive and Resistant High-Grade Serous Ovarian Cancer: Development and Applications of Experimental Systems

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