Title:	A novel detection technology for enrichment of rare and viable tumor cells in blood of patients with metastatic diseases.
Authors:	Wen-Tien Chen1,2, Qiang Zhao1, Wei Zeng1, Donghai Chen1, Stanley Zucker1, Stefan Madajewicz1, Hossein Zarrabi1, Daniel Baram1, Paul Richman1, Michael Pearl1, John Chen1, Michael Frohman1, Marc G. Golightly1, Martin Karpeh1, Huan Dong1,2, Kwan-nan Yeh1 & Yunyun Yeh1 (1) Department of Medicine, (3) Department of Obstetrics, Gynecology and Reproductive Medicine; (4) Department of Preventive Medicine; (5) Department of Pharmacology; (6) Department of Pathology; (7) Department of Surgeon, State University of New York, Stony Brook NY 11794; (2) Vitatex Inc., 25 Health Sciences Drive, Stony Brook, NY 11790.
Presented At:	International Conference on "Tumor Progression & Therapeutic Resistance", at the Hilton City Avenue in Philadelphia, Pennsylvania, USA.
Date Presented:	November 8th & 9th, 2004
Abstract
INTRODUCTION: It has become increasingly clear that the metastatic potential of a solid tumor correlates best with the presence of these cells in the systemic circulation. However, molecular analyses of these cells remains technically challenging because these emigrating tumor cells are few in number relative to the enormous numbers of normal blood cells and because many of the tumor cells found in peripheral blood are nonviable. METHODS: We have developed a novel cell separation technology based on preferential adhesion of tumor cells to cell adhesion matrix (CAM) coated surfaces. One million-fold enrichment of viable human tumor cells from the peripheral blood of healthy donors in model experiments, as well as in patients with various metastatic diseases can be achieved in a single step. Immunocytochemistry, flow cytometry and real-time RT-PCR using multiple epithelial/tumor associated cell lineage markers were used to validate the CAM-recovered cells from blood as circulating tumor cells. RESULTS: 20 mL of blood from 125 patients with metastatic cancer was collected and analyzed using the CAM technique. CAM recovered cells were viable based on Molecular Probes LIVE/DEAD Viability testing. Cells were then amplified 1,000 folds in culture for use in the molecular analysis of cancer progression and therapeutic responsiveness. CONCLUSION: Cell separation based on preferential adhesion of tumor cells to CAM coated surfaces is a rapid and reliable method of separating viable circulating tumor cells from blood in humans.