Observational Study
Copyright ©2014 Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Dec 7, 2014; 20(45): 17163-17170
Published online Dec 7, 2014. doi: 10.3748/wjg.v20.i45.17163
Circulating tumor cells in pancreatic cancer patients: Enrichment and cultivation
Vladimir Bobek, Robert Gurlich, Petra Eliasova, Katarina Kolostova
Vladimir Bobek, Katarina Kolostova, Department of Laboratory Genetics, University Hospital Kralovske Vinohrady, 10034 Prague, Czech Republic
Vladimir Bobek, Department of Histology and Embryology, Wroclaw Medical University, 50-367 Wrocław, Poland
Vladimir Bobek, 3rd Department of Surgery, First Faculty of Medicine Charles University in Prague and University Hospital Motol, 10034 Prague, Czech Republic
Robert Gurlich, Petra Eliasova, Department of Surgery, University Hospital Kralovske Vinohrady, 0034 Prague, Czech Republic
Author contributions: Bobek V and Kolostova K designed the study; Kolostova K and Eliasova P performed laboratory examination and collection of the data; Gurlich R collected the samples and analyzed clinical data; Bobek V and Kolostova K wrote the manuscripts; all authors co-authored the final text.
Supported by Research project “CTC in gastrointestinal cancer “awarded by League Against Cancer Prague, Czech Republic and grant of the Czech Ministry of Health, No. IGA NT14439-3/2013; and by the projects by Ministry of Helath Czech Republic, conceptual development of research organization, University Hospital Motol, Prague Czech Republic, No. 000 64203
Correspondence to: Vladimir Bobek MD, PhD, Department of Laboratory Genetics, University Hospital Kralovske Vinohrady, Srobarova 50, 10034 Prague, Czech Republic. vbobek@centrum.cz
Telephone: +42-26-716262 Fax: +42-26-716262
Received: February 17, 2014
Revised: May 20, 2014
Accepted: July 15, 2014
Published online: December 7, 2014

AIM: To investigate the feasibility of separation and cultivation of circulating tumor cells (CTCs) in pancreatic cancer (PaC) using a filtration device.

METHODS: In total, 24 PaC patients who were candidates for surgical treatment were enrolled into the study. Peripheral blood samples were collected before an indicated surgery. For each patient, approximately 8 mL of venous blood was drawn from the antecubital veins. A new size-based separation MetaCell® technology was used for enrichment and cultivation of CTCs in vitro. (Separated CTCs were cultured on a membrane in FBS enriched RPMI media and observed by inverted microscope. The cultured cells were analyzed by means of histochemistry and immunohistochemistry using the specific antibodies to identify the cell origin.

RESULTS: CTCs were detected in 16 patients (66.7%) of the 24 evaluable patients. The CTC positivity did not reflect the disease stage, tumor size, or lymph node involvement. The same percentage of CTC positivity was observed in the metastatic and non-metastatic patients (66.7% vs 66.7%). We report a successful isolation of CTCs in PaC patients capturing proliferating cells. The cells were captured by a capillary action driven size-based filtration approach that enabled cells cultures from the viable CTCs to be unaffected by any antibodies or lysing solutions. The captured cancer cells displayed plasticity which enabled some cells to invade the separating membrane. Further, the cancer cells in the “bottom fraction”, may represent a more invasive CTC-fraction. The CTCs were cultured in vitro for further downstream applications.

CONCLUSION: The presented size-based filtration method enables culture of CTCs in vitro for possible downstream applications.

Keywords: Pancreatic cancer, Circulating tumor cells, Biomarker, Cultivation

Core tip: Circulating tumor cells role in the process of pancreatic cancer dissemination should be studied in the context of the disease management. The ability to in vitro culture pancreatic circulating tumor cells (CTCs) could potentially help with the development of innovative treatments and diagnostic technologies. We presented simple size-based separation device for the isolation of viable CTCs. The isolation process is gentle allowing the subsequent CTC-cultivation in vitro and is antibody independent.