Isolation of circulating tumor cells from pancreatic cancer by automated filtration
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Nora Brychta1,*, Michael Drosch1,6,*, Christiane Driemel2, Johannes C. Fischer2, Rui P. Neves2, Irene Esposito3, Wolfram Knoefel2, Birte Möhlendick2, Claudia Hille1,5, Antje Stresemann1, Thomas Krahn1, Matthias U. Kassack4, Nikolas H. Stoecklein2 and Oliver von Ahsen1
1Bayer AG, Biomarker Research, 13353 Berlin, Germany
2Department of General, Visceral and Pediatric Surgery, Medical Faculty, University Hospital of the Heinrich-Heine-University Duesseldorf, 40225 Duesseldorf, Germany
3Institute of Pathology, Heinrich-Heine-University of Duesseldorf, 40225 Duesseldorf, Germany
4Institute of Pharmaceutical & Medicinal Chemistry, University of Duesseldorf, 40225 Duesseldorf, Germany
5Current/Present address: University Medical Center Hamburg-Eppendorf, Department of Tumor Biology, 20246 Hamburg, Germany
6Current/Present address: JPT Peptide Technologies GmbH, 12489 Berlin, Germany
*These authors have contributed equally to this work
Oliver von Ahsen, email: firstname.lastname@example.org
Nikolas H. Stoecklein, email: email@example.com
Keywords: pancreatic cancer, circulating tumor cells, KRAS, diagnostic leukapheresis, EMT
Received: March 24, 2017 Accepted: August 07, 2017 Published: September 16, 2017
It is now widely recognized that the isolation of circulating tumor cells based on cell surface markers might be hindered by variability in their protein expression. Especially in pancreatic cancer, isolation based only on EpCAM expression has produced very diverse results. Methods that are independent of surface markers and therefore independent of phenotypical changes in the circulating cells might increase CTC recovery also in pancreatic cancer. We compared an EpCAM-dependent (IsoFlux) and a size-dependent (automated Siemens Healthineers filtration device) isolation method for the enrichment of pancreatic cancer CTCs. The recovery rate of the filtration based approach is dramatically superior to the EpCAM-dependent approach especially for cells with low EpCAM-expression (filtration: 52%, EpCAM-dependent: 1%). As storage and shipment of clinical samples is important for centralized analyses, we also evaluated the use of frozen diagnostic leukapheresis (DLA) as source for isolating CTCs and subsequent genetic analysis such as KRAS mutation detection analysis. Using frozen DLA samples of pancreatic cancer patients we detected CTCs in 42% of the samples by automated filtration.
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