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

Kiki C Andree, Fikri Abali, Lisa Oomens, Fiona R Passanha, Joska J Broekmaat, Jaco Kraan, Pauline A J Mendelaar, Stefan Sleijfer, Leon W M M Terstappen

Int J Mol Sci. 2019 Jan 23;20(3):477.

PMID: 30678037

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP61926225	Ethidium homodimer		61926-22-5	Price