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Int J Mol Sci. 2019 Jan 23;20(3). pii: E477. doi: 10.3390/ijms20030477.

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

Author information

1
Medical Cell BioPhysics, University of Twente, Hallenweg 23, 7522 NH Enschede, The Netherlands. k.c.andree@utwente.nl.
2
Medical Cell BioPhysics, University of Twente, Hallenweg 23, 7522 NH Enschede, The Netherlands. f.abali@utwente.nl.
3
VyCAP B.V., Abraham Rademakerstraat 41, 7425 PG Deventer, The Netherlands. Lisa.oomens@vycap.com.
4
Medical Cell BioPhysics, University of Twente, Hallenweg 23, 7522 NH Enschede, The Netherlands. f.passanha@maastrichtuniversity.nl.
5
VyCAP B.V., Abraham Rademakerstraat 41, 7425 PG Deventer, The Netherlands. joska.broekmaat@vycap.com.
6
Department of Medical Oncology, Erasmus MC Cancer Institute and Cancer Genomics Netherlands, Dr. Molewaterplein 40, 3015 GD Rotterdam, the Netherlands. j.kraan@erasmusmc.nl.
7
Department of Medical Oncology, Erasmus MC Cancer Institute and Cancer Genomics Netherlands, Dr. Molewaterplein 40, 3015 GD Rotterdam, the Netherlands. p.mendelaar@erasmusmc.nl.
8
Department of Medical Oncology, Erasmus MC Cancer Institute and Cancer Genomics Netherlands, Dr. Molewaterplein 40, 3015 GD Rotterdam, the Netherlands. s.sleijfer@erasmusmc.nl.
9
Medical Cell BioPhysics, University of Twente, Hallenweg 23, 7522 NH Enschede, The Netherlands. l.w.m.m.terstappen@utwente.nl.

Abstract

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.

KEYWORDS:

CTC; breast cancer; prostate cancer; self-seeding microwells; single cell isolation

PMID:
30678037
PMCID:
PMC6387105
DOI:
10.3390/ijms20030477
[Indexed for MEDLINE]
Free PMC Article

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