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Open AccessArticle

Magnetic-Based Enrichment of Rare Cells from High Concentrated Blood Samples

1
Department of General, Visceral and Pediatric Surgery, University Hospital and Medical Faculty of the Heinrich-Heine University Düsseldorf, Moorenstr. 5, 40225 Düsseldorf, Germany
2
Institute for Transplantation Diagnostics and Cell Therapeutics, University Hospital and Medical Faculty of the Heinrich-Heine University Düsseldorf, 40225 Düsseldorf, Germany
3
Thermo Fisher Scientific, Postfach 200152, Frankfurter Str. 129B, 64293 Darmstadt, Germany
4
Thermo Fisher Scientific, 3 Fountain Drive, Inchinnan, Renfrew PA4 9RF, UK
*
Authors to whom correspondence should be addressed.
Cancers 2020, 12(4), 933; https://doi.org/10.3390/cancers12040933
Received: 2 March 2020 / Revised: 30 March 2020 / Accepted: 8 April 2020 / Published: 10 April 2020
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
Here, we tested two magnetic-bead based systems for the enrichment and detection of rare tumor cells in concentrated blood products. For that, the defined numbers of cells from three pancreatic cancer cell lines were spiked in 108 peripheral blood mononuclear cells (PBMNCs) concentrated in 1 mL, mimicking diagnostic leukapheresis (DLA) samples, and samples were processed for circulating tumor cells (CTC) enrichment with the IsoFlux or the KingFisher systems, using different types of magnetic beads from the respective technology providers. Beads were conjugated with different anti-EpCAM and MUC-1 antibodies. Recovered cells were enumerated and documented by fluorescent microscopy. For the IsoFlux system, best performance was obtained with IsoFlux CTC enrichment kit, but these beads compromised the subsequent immunofluorescence staining. For the KingFisher system, best recoveries were obtained using Dynabeads Biotin Binder beads. These beads also allowed one to capture CTCs with different antibodies and the subsequent immunofluorescence staining. KingFisher instrument allowed a single and streamlined protocol for the enrichment and staining of CTCs that further prevented cell loss at the enrichment/staining interface. Both IsoFlux and KingFisher systems allowed the enrichment of cell line cells from the mimicked-DLA samples. However, in this particular experimental setting, the recovery rates obtained with the KingFisher system were globally higher, the system was more cost-effective, and it allowed higher throughput. View Full-Text
Keywords: circulating tumor cells; immunomagnetic enrichment; concentrated blood products; diagnostic leukapheresis; IsoFlux; KingFisher circulating tumor cells; immunomagnetic enrichment; concentrated blood products; diagnostic leukapheresis; IsoFlux; KingFisher
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MDPI and ACS Style

Wu, J.; Raba, K.; Guglielmi, R.; Behrens, B.; Van Dalum, G.; Flügen, G.; Koch, A.; Patel, S.; Knoefel, W.T.; Stoecklein, N.H.; Neves, R.P.L. Magnetic-Based Enrichment of Rare Cells from High Concentrated Blood Samples. Cancers 2020, 12, 933.

AMA Style

Wu J, Raba K, Guglielmi R, Behrens B, Van Dalum G, Flügen G, Koch A, Patel S, Knoefel WT, Stoecklein NH, Neves RPL. Magnetic-Based Enrichment of Rare Cells from High Concentrated Blood Samples. Cancers. 2020; 12(4):933.

Chicago/Turabian Style

Wu, Junhao; Raba, Katharina; Guglielmi, Rosa; Behrens, Bianca; Van Dalum, Guus; Flügen, Georg; Koch, Andreas; Patel, Suraj; Knoefel, Wolfram T.; Stoecklein, Nikolas H.; Neves, Rui P.L. 2020. "Magnetic-Based Enrichment of Rare Cells from High Concentrated Blood Samples" Cancers 12, no. 4: 933.

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