Published online Dec 7, 2014. doi: 10.3748/wjg.v20.i45.17163
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.
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.