Establishment, Characterization and Downstream Application of Primary Ovarian Cancer Cells Derived from Solid Tumors

Thanasak Sueblinvong, Rahel G Ghebre, Yoshie Iizuka, Stefan E. Pambuccian, Rachel I Vogel, Amy P Skubitz, Martina Bazzaro

Research output: Contribution to journalArticle

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Abstract

Ovarian cancer is the deadliest of the gynecological diseases and the fifth cause of cancer death among American women. This is mainly due to the lack of prognostic tools capable of detecting early stages of ovarian cancer and to the high rate of resistance to the current chemotherapeutic regimens. In this scenario the overall 5-year survival rate for ovarian cancer patients diagnosed at late stage is less than 25%. Abnormalities associated with the malignant phenotype and the mechanisms of tumor progression are not clearly understood. In vitro studies are necessary, yet have been hampered due to the limitations accompanied with the use of ovarian cancer cell lines and the heterogeneity of the ovarian cancer cell population derived from ascites fluids. In this study we present a simple, rapid and reproducible method for the isolation and characterization of ovarian cancer cells from solid tumor tissue and show that enzymatic digestion for 30 minutes with dispase II results in the most effective recovery of viable epithelial ovarian cancer (EOC) cells. The resulting cancer (EOC) cell preparations demonstrate a significant yield, high levels of viability and are fibroblast-free. They grow for up to six passages and retain the capacity of forming spheroids-like structures in agarose. In addition, they can be genetically manipulated and used for drug screening, thus rendering them highly suitable for downstream applications. Notably, isolation of ovarian cancer cells from solid specimens using this method has the advantage of allowing for isolation of cancer cells from early stages of ovarian cancer as well as obtaining cells from defined either primary and/or metastatic ovarian cancer sites. Thus, these cells are highly suitable for investigations aimed at understanding ovarian cancer.

Original languageEnglish (US)
Article numbere50519
JournalPLoS One
Volume7
Issue number11
DOIs
StatePublished - Nov 30 2012

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ovarian neoplasms
Ovarian Neoplasms
Tumors
Cells
neoplasms
Neoplasms
Fibroblasts
Sepharose
Erythema Infectiosum
neoplasm cells
Screening
Preclinical Drug Evaluations
Tissue
Cell Separation
Recovery
Fluids
Ascites
ascites
isolation techniques
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Establishment, Characterization and Downstream Application of Primary Ovarian Cancer Cells Derived from Solid Tumors. / Sueblinvong, Thanasak; Ghebre, Rahel G; Iizuka, Yoshie; Pambuccian, Stefan E.; Vogel, Rachel I; Skubitz, Amy P; Bazzaro, Martina.

In: PLoS One, Vol. 7, No. 11, e50519, 30.11.2012.

Research output: Contribution to journalArticle

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