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Primary culture of ovarian surface epithelial cells and ascites-derived ovarian cancer cells from patients

Nature Protocols volume 1, pages 2643–2649 (2006)Cite this article

A Corrigendum to this article was published on 27 August 2015

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Abstract

Our laboratory has refined the technique for isolating primary cultures of normal human ovarian surface epithelial (OSE) cells by combining two different protocols involving the enzymatic and mechanical removal of OSE cells from ovarian biopsies. A simple protocol of obtaining primary epithelial ovarian cancer (EOC) cells from the ascites fluid removed from patients with high-grade ovarian cancer is also described. These methods allow for the direct application of many molecular and cellular analyses of normal versus cancer cells isolated freshly from patients, with the added potential for retrospective analyses of archived cells and tissues. Thus, we have included optional steps for the immediate preparation of ascites-derived EOC cells to be used for subsequent cytological analyses. Initial isolation of OSE or EOC cells can be completed in 1 h, and primary cells are further expanded in culture for several weeks.

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Figure 1: Primary cultures of normal human OSE cells.
Figure 2: Primary cultures of human EOC cells from patient ascites.
Figure 3: Downstream applications using primary human EOC cells.

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  • 05 August 2015

    In the version of this article initially published, in the Reagent Setup section, it was stated that 50% v/v Percoll should be made by diluting Percoll in 10× PBS at a 1:1 v/v ratio. Percoll should, in fact, be diluted in 2× PBS at a 1:1 ratio. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

This work was supported by the National Cancer Institute of Canada with funds from the Canadian Cancer Society Grant No. 15303. T.G.S. is supported by a fellowship from the NCIC with funds from the Terry Fox Foundation. B.T. is supported by a fellowship from the Nova Scotia Health Research Foundation. M.W.N. is a Canadian Cancer Society Scientist of the NCIC.

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  1. Department of Pharmacology, Dalhousie University, 5850 College Street, Halifax, B3H 1X5, Nova Scotia, Canada

    Trevor G Shepherd, Brigitte L Thériault, Elizabeth J Campbell & Mark W Nachtigal

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Correspondence to Mark W Nachtigal.

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Shepherd, T., Thériault, B., Campbell, E. et al. Primary culture of ovarian surface epithelial cells and ascites-derived ovarian cancer cells from patients. Nat Protoc 1, 2643–2649 (2006). https://doi.org/10.1038/nprot.2006.328

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