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Article

Pre-Analytical and Analytical Variables of Label-Independent Enrichment and Automated Detection of Circulating Tumor Cells in Cancer Patients

1
Department of Tumor Biology, Center of Experimental Medicine, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany
2
Department of Oncology, Hematology and Bone Marrow Transplantation with section Pneumology, Hubertus Wald Tumorzentrum, University Comprehensive Cancer Center Hamburg, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany
3
Department of Gynecology and Gynecologic Oncology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany
4
Department of Gynecology, Asklepios Clinic Barmbek, 22307 Hamburg, Germany
5
Department of Gynecology and Obstetrics, Marienkrankenhaus Hamburg, 22087 Hamburg, Germany
6
Department of Transfusion Medicine, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Cancers 2020, 12(2), 442; https://doi.org/10.3390/cancers12020442
Received: 6 January 2020 / Revised: 10 February 2020 / Accepted: 11 February 2020 / Published: 13 February 2020
(This article belongs to the Special Issue Liquid Biopsy in Cancer)
Circulating tumor cells (CTCs) are promising tools for risk prediction and the monitoring of response to therapy in cancer patients. Within the EU/IMI CANCER-ID consortium, we validated CTC enrichment systems for future inclusion into clinical trials. Due to the known heterogeneity of markers expressed on CTCs, we tested the Parsortix® system (ANGLE plc) which enables label-independent CTC enrichment from whole blood based on increased size and deformability of these tumor cells compared to leukocytes. We performed extensive comparisons both with spiked-in blood models (i.e., MDA-MB-468 tumor cell line cells spiked at very low concentration into blood from healthy donors) and validated the protocol on actual clinical samples from breast, lung, and gastrointestinal cancer patients to define optimal conditions for CTC enrichment. Multiple parameters including cassette gap, separation pressure, and cell fixatives were compared in parallel. Also, the compatibility of blood collection tubes with whole genome amplification of isolated tumor cells was demonstrated and we furthermore established a workflow for semi-automated CTC detection using a quantitative cell imager. The established workflow will contribute to supporting the use of size-based CTC enrichment platforms in clinical trials testing the clinical validity and utility of CTCs for personalized medicine. View Full-Text
Keywords: circulating tumor cells; CTC; enrichment; Parsortix; cancer metastasis circulating tumor cells; CTC; enrichment; Parsortix; cancer metastasis
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MDPI and ACS Style

Koch, C.; Joosse, S.A.; Schneegans, S.; Wilken, O.J.W.; Janning, M.; Loreth, D.; Müller, V.; Prieske, K.; Banys-Paluchowski, M.; Horst, L.J.; Loges, S.; Peine, S.; Wikman, H.; Gorges, T.M.; Pantel, K. Pre-Analytical and Analytical Variables of Label-Independent Enrichment and Automated Detection of Circulating Tumor Cells in Cancer Patients. Cancers 2020, 12, 442. https://doi.org/10.3390/cancers12020442

AMA Style

Koch C, Joosse SA, Schneegans S, Wilken OJW, Janning M, Loreth D, Müller V, Prieske K, Banys-Paluchowski M, Horst LJ, Loges S, Peine S, Wikman H, Gorges TM, Pantel K. Pre-Analytical and Analytical Variables of Label-Independent Enrichment and Automated Detection of Circulating Tumor Cells in Cancer Patients. Cancers. 2020; 12(2):442. https://doi.org/10.3390/cancers12020442

Chicago/Turabian Style

Koch, Claudia, Simon A. Joosse, Svenja Schneegans, Okka J.W. Wilken, Melanie Janning, Desiree Loreth, Volkmar Müller, Katharina Prieske, Malgorzata Banys-Paluchowski, Ludwig J. Horst, Sonja Loges, Sven Peine, Harriet Wikman, Tobias M. Gorges, and Klaus Pantel. 2020. "Pre-Analytical and Analytical Variables of Label-Independent Enrichment and Automated Detection of Circulating Tumor Cells in Cancer Patients" Cancers 12, no. 2: 442. https://doi.org/10.3390/cancers12020442

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