Senologie - Zeitschrift für Mammadiagnostik und -therapie 2015; 12 - A82
DOI: 10.1055/s-0035-1550523

Detection of EpCAM-negative circulating tumor cells by using VyCAP filters technology

R Lampignano 1, H Schneck 1, M Neumann 1, D Köhler 1, L Terstappen 2, D Niederacher 1, T Fehm 1, H Neubauer 1
  • 1Department of Obstetrics and Gynaecology, Medical Faculty, University Hospital, Heinrich Heine University, Duesseldorf, Deutschland, Düsseldorf, Deutschland
  • 2Universiteit Twente, Niederlande, Twente, Niederlande

Einleitung/Zielsetzung: Circulating tumor cells (CTCs) are epithelial cells identified in the peripheral blood stream of epithelial cancer and are believed to be their metastatic precursors. Their exact composition is unknown. It has been discovered that they possess similar characteristics as their primary tumor but also show different phenotypes ranging from epithelial to mesenchymal. Detection of CTCs in peripheral blood provides prognostic information and might also represent an alternative to the invasive bone marrow biopsy.

Nowadays CTCs are mostly detected and enriched by EpCAM-based enrichment systems. However, they cannot detect CTCs that have downregulated epithelial markers e.g. in the process called 'epithelial-mesenchymal transition'.

In a project funded by the EU ('CTCtrap: Circulating Tumor Cells TheRapeutic APheresis: a novel biotechnology enabling personalized therapy for all cancer patients „ Grant agreement no: 305341) a protocol was established to detect EpCAM-negative CTCs by filtration using the VyCap-Filters technology.

Material & Methoden: 35 – 40mL volume of samples are filtered through 5 µm pore sized silicon nitride filters with a constant pressure of 100mbar. After enrichment, cells are stained for cytokeratins and CD45 to identify cells of epithelial origin. Filters are automatically scanned with an automated fluorescence microscope CellCelector™. We established a protocol for automated scanning of the VyCap filters comprising screening for DAPI and FITC positivity and CD45 negativity with 20x magnification.

Ergebnisse: We detected 40 EpCAM-negative CTCs from 7 blood samples.

Zusammenfassung: Further molecular analysis are needed to proof their malignant origin and to compare the biological properties of EpCAM-positive and EpCAM-negative CTCs identified in same blood samples.