Cancer Diagnosis Program | Biorepositories and Biospecimen Research Branch

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Microtube device for selectin-mediated capture of viable circulating tumor cells from blood.

Author(s): Hughes AD, Mattison J, Western LT, Powderly JD, Greene BT, King MR

Publication: Clin Chem, 2012, Vol. 58, Page 846-53

PubMed ID: 22344286 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of cell capture method on the number of viable circulating tumor cells (CTCs) isolated from buffy coat specimens from cancer patients and healthy volunteers.  Blood (7.5 ml) was collected into EDTA or BD Vacutainer cell preparation tubes from twelve patients diagnosed with stage IV cancer of the breast (6 patients), prostate (3 patients), lung (2 patients), or ovary (1 patient).  Blood (7.5 ml) was also collected in BD Vacutainer tubes from eight healthy donors.  An additional 10 ml of blood was collected into CellSave tubes and processed within 96 h. Buffy coat was isolated according to CellSave protocols or by centrifugation of plasma at 1500 x g for 20 min at ambient temperature, frozen in freezing medium, and then stored in liquid nitrogen until culture. A subset of specimens from cancer patients were collected in BD Vacutainer EDTA tubes and shipped overnight at ambient temperatures before isolation of buffy coat and analysis, while others were shipped overnight on dry ice as buffy coat frozen in freezing medium. Specimens from healthy volunteers underwent buffy coat isolation immediately and buffy coat was left at room temperature for 24 h before cell capture and analysis. Primary cells were perfused through smooth tubes or halloysite-coated tubes that were identically coated with anti-EpCAM or anti-PSMA antibodies. Captured cells were cultured for five days, stained with epithelial markers (anti-EpCAM or anti-PSMA) and DAPI, and visualized with fluorescent microscopy for detection of CTCs.

   Summary of Findings:

   The mean purity of captured CTCs was greater when a halloysite nanotube coating was applied to the MicroRenathane microtube (66.0%) compared to use of the smooth microtube (37.2%, p=0.004). Significantly fewer CTCs were captured from blood collected from healthy donors compared to blood from patients diagnosed with metastatic cancer for both the smooth and nanotube-coated tubes (0-1.9 vs. 50-200 cells); however, blood from three of the eight healthy donors tested positive with either nanotube. Comparatively, the CellSearch CTC kit captured fewer than two CTCs per 3.75 mL in blood from eight of 12 patients diagnosed with metastatic cancer, demonstrating a substantial discord with the microtube devices which captured 172 viable CTCs (smooth device average). Notably, blood from three of the eight healthy donors also tested positive by CellSearch. The potential influence of collection tube type was not investigated.

   Biospecimens

   • Bodily Fluid - Blood
   • Bodily Fluid - Buffy Coat

   Preservative Types

   • Frozen
   • None (Fresh)
   • Other Preservative

   Diagnoses:

   • Normal
   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   Cell count/volume	Fluorescent microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Aliquots and Components	Cell capture method	MicroRenathane microtube (smooth) MicroRenathane microtube (halloysite nanotube coating) CellSearch CTC kit
   Preaquisition	Diagnosis/ patient condition	Healthy Breast cancer Prostate cancer Lung cancer Ovarian cancer

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