Geburtshilfe Frauenheilkd 2012; 72 - A38
DOI: 10.1055/s-0032-1309231

Identification of Cancer Stem Cells in Primary Ovarian Carcinoma

IKC Wasle 1, M Boesch 1, S Sopper 1, D Reimer 2, D Wolf 3, AG Zeimet 2
  • 1Department Tumorimmunology, Innsbruck University Hospital, Innsbruck
  • 2Department of Obstetrics and Gynecology, Innsbruck University Hospital, Innsbruck
  • 3Department Internal Medicine III/Hemato-Oncology, Bonn University Hospital, Bonn

Fragestellung:

In a wide range of tumor entities cancer stem cells were identified as subpopulation with a higher tumorigenic potential than the bulk of tumor cells. The aim of this study is to find cancer stem cells in the primary ovarian carcinoma and to study differences in molecular and functional biology.

Methode:

Tumor samples were taken from primary ovarian carcinoma and a protocol based on collagenase digestion to disrupt the tissue into single cell suspensions was developed using the Miltenyi Gentle MACS Dissociator®. Single cell suspensions were analyzed for viability and short term cell culture were established. By using 9-color flow cytometry and specific markers, e.g. Vimentin, EpCAM, Cytokeratin, isolated cells were screened for putative cancer stem cells with surface markers like CD24, CD44, CD117, CD133 and L1CAM. Besides we analyzed the cells for efflux of Dye Cycle Violet resulting in a side-population, thought to be characteristic for stem cells.

Ergebnisse:

Development and validation of the protocol for tumor tissue digestion resulted in using the digestion enzymes Miltenyi Tumor Dissociation Kit® and a digestion time of 45 minutes. With this combination we obtained single cell suspensions of high viability. Yield ranged between 9–13 million cells/gram tumor. First FACS analyses showed distinct populations of CD24, CD44, EpCAM and L1CAM positive cells.

Schlussfolgerung:

Using the established methods, we will compare the subpopulations isolated by FACS sorting, for expression patterns and functional properties of stem cells by using tumor tissue from 25 patients (15 after primary/10 after cytotoxic debulking surgery). Once populations with stem cell characteristics are defined, they can be used to validate therapeutic approaches aimed at eliminating cancer stem cells.