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Isolation of Mononuclear Cell Populations from Ovarian Carcinoma Ascites   

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Original research article

A brief version of this protocol appeared in:
Cancer Research
Aug 2016

Abstract

Ovarian cancer is one of the most fatal tumors in women. Due to a lack of symptoms and adequate screening methods, patients are diagnosed at advanced stages with extensive tumor burden (Jelovac and Armstrong, 2011). Interestingly, ovarian cancer metastasis is generally found within the peritoneal cavity rather than other tissues (Lengyel, 2010; Tan et al., 2006). The reason behind this tissue tropism of the peritoneal cavity remains elusive. A prominent feature of this selectivity is ascites, the accumulation of fluid within the peritoneal cavity, containing, amongst others, immune cells, tumor cells and various soluble factors that can be involved in the progression of ovarian cancer (Kipps et al., 2013). The protocol described here is used to isolate mononuclear cells from ascites to study the functionality of the immune system within the peritoneal cavity.

Keywords: Ovarian cancer, Ascites, Fluorescent activated cell sorting, Mononuclear cells, Dendritic cells, Monocytes, Myeloid-derived suppressive cells, BDCA1+CD14+ cells

Background

Gradient centrifugation using LymphoprepTM is a standard protocol to isolate peripheral blood mononuclear cells (PBMCs). We slightly adjusted the protocol, regarding the sample preparation and amount of washing steps, in order to isolate mononuclear cells from ascites.

Copyright: © 2017 The Authors; exclusive licensee Bio-protocol LLC.
How to cite: Wefers, C., Bakdash, G., Moreno Martin, M., Duiveman-de Boer, T., Torensma, R., Massuger, L. F. and de Vries, I. M. (2017). Isolation of Mononuclear Cell Populations from Ovarian Carcinoma Ascites. Bio-protocol 7(7): e2219. DOI: 10.21769/BioProtoc.2219.
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