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Int. J. Mol. Sci. 2019, 20(3), 477;

Self-Seeding Microwells to Isolate and Assess the Viability of Single Circulating Tumor Cells

Medical Cell BioPhysics, University of Twente, Hallenweg 23, 7522 NH Enschede, The Netherlands
VyCAP B.V., Abraham Rademakerstraat 41, 7425 PG Deventer, The Netherlands
Department of Medical Oncology, Erasmus MC Cancer Institute and Cancer Genomics Netherlands, Dr. Molewaterplein 40, 3015 GD Rotterdam, the Netherlands
Author to whom correspondence should be addressed.
Received: 2 December 2018 / Revised: 9 January 2019 / Accepted: 17 January 2019 / Published: 23 January 2019
(This article belongs to the Special Issue Single Cell Technology)
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The availability of viable tumor cells could significantly improve the disease management of cancer patients. Here we developed and evaluated a method using self-seeding microwells to obtain single circulating tumor cells (CTC) and assess their potential to expand. Conditions were optimized using cells from the breast cancer cell line MCF-7 and blood from healthy volunteers collected in EDTA blood collection tubes. 43% of the MCF-7 cells (nucleus+, Ethidium homodimer-1-, Calcein AM+, α-EpCAM+, α-CD45-) spiked into 7.5 mL of blood could be recovered with 67% viability and these could be further expanded. The same procedure tested in metastatic breast and prostate cancer patients resulted in a CTC recovery of only 0–5% as compared with CTC counts obtained with the CellSearch® system. Viability of the detected CTC ranged from 0–36%. Cell losses could be mainly contributed to the smaller size and greater flexibility of CTC as compared to cultured cells from cell lines and loss during leukocyte depletion prior to cell seeding. Although CTC losses can be reduced by fixation, to obtain viable CTC no fixatives can be used and pore size in the bottom of microwells will need to be reduced, filtration conditions adapted and pre-enrichment improved to reduce CTC losses. View Full-Text
Keywords: CTC; self-seeding microwells; single cell isolation; breast cancer; prostate cancer CTC; self-seeding microwells; single cell isolation; breast cancer; prostate cancer

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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Andree, K.C.; Abali, F.; Oomens, L.; Passanha, F.R.; Broekmaat, J.J.; Kraan, J.; Mendelaar, P.A.; Sleijfer, S.; Terstappen, L.W. Self-Seeding Microwells to Isolate and Assess the Viability of Single Circulating Tumor Cells. Int. J. Mol. Sci. 2019, 20, 477.

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Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
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