title: Filtration parameters influencing circulating tumor cell enrichment from whole blood.
abstract: Filtration can achieve circulating tumor cell (CTC) enrichment from blood. Key parameters such as flow-rate, applied pressure, and fixation, vary largely between assays and their influence is not well understood. Here, we used a filtration system, to monitor these parameters and determine their relationships. Whole blood, or its components, with and without spiked tumor cells were filtered through track-etched filters. We characterize cells passing through filter pores by their apparent viscosity; the viscosity of a fluid that would pass with the same flow. We measured a ratio of 5·10(4)∶10(2)∶1 for the apparent viscosities of 15 µm diameter MDA-231 cells, 10 µm white cells and 90 fl red cells passing through a 5 µm pore. Fixation increases the pressure needed to pass cells through 8 µm pores 25-fold and halves the recovery of spiked tumor cells. Filtration should be performed on unfixed samples at a pressure of ∼10 mbar for a 1 cm(2) track-etched filter with 5 µm pores. At this pressure MDA-231 cells move through the filter in 1 hour. If fixation is needed for sample preservation, a gentle fixative should be selected. The difference in apparent viscosity between CTC and blood cells is key in optimizing recovery of CTC.
authors: Medical Cell BioPhysics, MIRA Institute, University of Twente, Enschede, The Netherlands.
authors: Frank A W
authors: FA
authors: Coumans
authors: Guus
authors: G
authors: van Dalum
authors: Markus
authors: M
authors: Beck
authors: Leon W M M
authors: LW
authors: Terstappen
mesh_terms: D000328:Adult; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods / Q000706:statistics & numerical data*; D004912:Erythrocytes / Q000166:cytology; D005374:Filtration / Q000379:methods / Q000706:statistics & numerical data*; D005404:Fixatives; D018056:Hemorheology; D006801:Humans; D007962:Leukocytes / Q000166:cytology; D008875:Middle Aged; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D011312:Pressure; D014783:Viscosity
chemical_list: D005404:Fixatives
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