Circulating tumor cells (CTCs) are important targets for treatment and critical surrogate markers when evaluating cancer prognosis and therapeutic response. A sensitive methodology for detecting CTCs in gastric cancer (GC) patients is needed. In this study we demonstrate a device for enrichment and cultivation of CTCs. In total, 22 patients with GC, all candidates for surgery, were enrolled in the study. Peripheral blood samples were collected before surgery, and patients were re-evaluated within operation and divided into two groups: resectable and non-resectable GC. A new size-based separation test for enrichment and cultivation of CTCs was used (MetaCell(®)). In addition to cytomorphological analysis, gene expression of tumor associated genes (Cytokeratin-18, Cytokeratin-19, Cytokeratin-20, Cytokeratin-7, EPCAM, MUC1, HER2, EGFR) and of leukocyte markers (e.g. CD45, CD68) was tested in enriched CTC fractions. CTCs were detected in 59 % of the patients studied (n = 13/22). CTCs were detected in seven patients of the resection group (7/10, 70 %) and six of the non-resectable group (6/12, 50 %). Enrichment of the viable CTCs allowed subsequent successful cultivation in vitro. The cytomorphological characterization of the CTCs was a prerequisite of random gene expression testing in CTC-positive samples. In CTC-positive samples gene expression of cytokeratin 18 and 19 was elevated in comparison to the whole blood gene expression analysis. CTCs were found to be present in both resectable and non-resectable gastric cancer patients. The size-based separation platform for CTCs may be used for in vitro cultivation, as well as in subsequent molecular analysis if desired. The sensitivity of CTC-detection could be enhanced by the combination of cytomorphological and molecular analysis.

3rd Department of Surgery 1st Faculty of Medicine Charles University Prague and University Hospital Motol Prague Czech Republic

Department of Gastrointestinal and General Surgery Wroclaw Medical University ul M Curie Skłodowskiej 66 50 369 Wrocław Poland

Department of Histology and Embryology Wroclaw Medical University Chalubinskiego 6a 50 368 Wroclaw Poland

Department of Laboratory Genetics University Hospital Kralovske Vinohrady Srobarova 50 100 34 Prague Czech Republic

Department of Surgery 3rd Faculty of Medicine Charles University Prague and University Hospital Kralovske Vinohrady Srobarova 50 Prague Czech Republic

Division of Oncological Surgery Department of Oncology Wroclaw Medical University Plac Hirszfelda 12 53 413 Wrocław Poland

Lower Silesian Oncology Centre Plac Hirszfelda 12 53 413 Wrocław Poland

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