title: Circulating tumor cells in pancreatic cancer patients: enrichment and cultivation.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  The presented size-based filtration method enables culture of CTCs in vitro for possible downstream applications.
authors.affiliation: Vladimir Bobek, Katarina Kolostova, Department of Laboratory Genetics, University Hospital Kralovske Vinohrady, 10034 Prague, Czech Republic.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Bobek
authors.affiliation: Vladimir Bobek, Katarina Kolostova, Department of Laboratory Genetics, University Hospital Kralovske Vinohrady, 10034 Prague, Czech Republic.
authors.forename: Robert
authors.initials: R
authors.lastname: Gurlich
authors.affiliation: Vladimir Bobek, Katarina Kolostova, Department of Laboratory Genetics, University Hospital Kralovske Vinohrady, 10034 Prague, Czech Republic.
authors.forename: Petra
authors.initials: P
authors.lastname: Eliasova
authors.affiliation: Vladimir Bobek, Katarina Kolostova, Department of Laboratory Genetics, University Hospital Kralovske Vinohrady, 10034 Prague, Czech Republic.
authors.forename: Katarina
authors.initials: K
authors.lastname: Kolostova
mesh_terms: D014408:Biomarkers, Tumor / Q000378:metabolism; D002465:Cell Movement; D049109:Cell Proliferation; D002469:Cell Separation / Q000295:instrumentation*; D005240:Feasibility Studies; D005374:Filtration / Q000295:instrumentation*; D006801:Humans; D008567:Membranes, Artificial; D009361:Neoplasm Invasiveness; D009360:Neoplastic Cells, Circulating / Q000378:metabolism / Q000473:pathology*; D010190:Pancreatic Neoplasms / Q000097:blood / Q000378:metabolism / Q000473:pathology*; D010641:Phenotype; D011076:Polycarboxylate Cement; D016062:Porosity; D014407:Tumor Cells, Cultured
chemical_list: D014408:Biomarkers, Tumor; D008567:Membranes, Artificial; D011076:Polycarboxylate Cement; C005506:polycarbonate
keywords: Biomarker; Circulating tumor cells; Cultivation; Pancreatic cancer