As invited speaker, Prof. Paterlini-Bréchot will attend the Oncotrans 2017 Congress with a presentation on the 2nd of February 2017 at 15:40 am entitled “Technical Insights into highly sensitive isolation and molecular characterization of circulating tumor cells for early detection of tumor invasion.”
An unsolved technical issue in the Circulating Tumor Cells (CTC) field is how to obtain highly sensitive and unbiased collection of these fragile and heterogeneous cells, in both live and fixed form, for their molecular study when they are extremely rare, thus at the beginning of the invasion process.
Material and Methods
We have explored the potential of ISET® (Isolation by SizE of Tumor Cells), an open platform developed for marker-independent isolation of tumor cells from blood, to collect intact fixed and live cancer cells by using spiking analyses with extremely low numbers of fluorescent cultured cells. Then we have assessed the feasibility of Next Generation Sequencing (NGS) analyses targeted to single cells enriched by ISET using the hotspot cancer panel v2 on the Ion Torrent platform. This panel can detect 6893 possible COSMIC mutations over 50 oncogenes and tumor suppressor genes.
Results and Discussion
Our results consistently show the feasibility of isolating fixed and live tumor cells with a sensitivity close to one cancer cell per 10 mL of blood. We also demonstrate the feasibility of NGS analysis of single live cells enriched by ISET® with Ion Torrent Plateform. Our data provide evidence that the analysis of three single cells enriched by ISET® is sufficient to reliably identify the mutated allele frequency in the target cell population.
These technical improvements should help the study of circulating cancer cells at the early steps of tumor invasion.
Further information here: http://oncotrans.fr/