Abstract

Analysis of Single Cells Released by a Detector Designed for in vivo Catching of Circulating Tumor Cells

Peter Sedlmayr, Associate Professor, Medical University of Graz

Detection of rare CTCs remains a challenge. The CellCollector^TM, a certified medical device allows for in vivo enrichment of CTC. This is a wire coated with anti-EPCAM antibodies which is introduced into the cubital vein for 30 min. More than just providing an indication for the patients’ prognosis based on the cell concentration, molecular analysis of CTC would provide more information for personalized treatment of cancer patients. In this study we therefore evaluated a new version of detector, namely the Catch and Release (C&R) detector (which is not yet certified for in vivo application), allowing for detachment and recovery of isolated cells. The C&R detector was loaded with cultured cells of carcinoma cell lines and forwarded to cell detachment. Detached cells were manipulated for single-cell whole genome amplification, array comparative genome hybridization (array-CGH) and targeted next-generation sequencing (NGS). 93% of cells attached to the C&R detector were successfully detached. Array-CGH profiles yielded high concordance between single cells recovered from C&R device and unamplified gDNA. NGS yielded concordant detection of five frequently reported SNPs among single cells and unamplified gDNA. DNA quality was not altered during the “C&R” procedure. This renders the C&R detector valuable for single CTC genomic analysis.