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

Kiki Andree, Fikri Abali, Lisa Oomens, Fiona Passanha, Joska Broekmaat, Jaco Kraan, Pauline Mendelaar, Stefan Sleijfer, Leon Terstappen (Corresponding Author)

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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.
Original languageEnglish
Article number477
JournalInternational journal of molecular sciences
Volume20
Issue number3
DOIs
Publication statusPublished - 23 Jan 2019

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Circulating Neoplastic Cells
inoculation
viability
Tumors
tumors
Cells
cultured cells
blood
Blood
cancer
cells
breast
Breast Neoplasms
Cell Line
Fixatives
leukocytes
MCF-7 Cells
Disease Management
ethylenediaminetetraacetic acids
Cell Nucleus

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title = "Self-Seeding Microwells to Isolate and Assess the Viability of Single Circulating Tumor Cells",
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{\circledR} 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.",
author = "Kiki Andree and Fikri Abali and Lisa Oomens and Fiona Passanha and Joska Broekmaat and Jaco Kraan and Pauline Mendelaar and Stefan Sleijfer and Leon Terstappen",
year = "2019",
month = "1",
day = "23",
doi = "10.3390/ijms20030477",
language = "English",
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Self-Seeding Microwells to Isolate and Assess the Viability of Single Circulating Tumor Cells. / Andree, Kiki; Abali, Fikri; Oomens, Lisa; Passanha, Fiona; Broekmaat, Joska; Kraan, Jaco; Mendelaar, Pauline; Sleijfer, Stefan; Terstappen, Leon (Corresponding Author).

In: International journal of molecular sciences, Vol. 20, No. 3, 477, 23.01.2019.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Self-Seeding Microwells to Isolate and Assess the Viability of Single Circulating Tumor Cells

AU - Andree, Kiki

AU - Abali, Fikri

AU - Oomens, Lisa

AU - Passanha, Fiona

AU - Broekmaat, Joska

AU - Kraan, Jaco

AU - Mendelaar, Pauline

AU - Sleijfer, Stefan

AU - Terstappen, Leon

PY - 2019/1/23

Y1 - 2019/1/23

N2 - 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.

AB - 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.

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