Self-Seeding Microwells to Isolate and Assess the Viability of Single Circulating Tumor Cells
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 ce...
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| Format: | Article |
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MDPI AG
2019-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/20/3/477 |
| _version_ | 1811197162097213440 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Kiki C. Andree |
| collection | DOAJ |
| description | 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<sup>®</sup> 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. |
| first_indexed | 2024-04-12T01:11:05Z |
| format | Article |
| id | doaj.art-6ed475e5ef174324bd2543c3a7c82bad |
| institution | Directory Open Access Journal |
| issn | 1422-0067 |
| language | English |
| last_indexed | 2024-04-12T01:11:05Z |
| publishDate | 2019-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-6ed475e5ef174324bd2543c3a7c82bad2022-12-22T03:54:06ZengMDPI AGInternational Journal of Molecular Sciences1422-00672019-01-0120347710.3390/ijms20030477ijms20030477Self-Seeding Microwells to Isolate and Assess the Viability of Single Circulating Tumor CellsKiki C. Andree0Fikri Abali1Lisa Oomens2Fiona R. Passanha3Joska J. Broekmaat4Jaco Kraan5Pauline A.J. Mendelaar6Stefan Sleijfer7Leon W.M.M. Terstappen8Medical Cell BioPhysics, University of Twente, Hallenweg 23, 7522 NH Enschede, The NetherlandsMedical Cell BioPhysics, University of Twente, Hallenweg 23, 7522 NH Enschede, The NetherlandsVyCAP B.V., Abraham Rademakerstraat 41, 7425 PG Deventer, The NetherlandsMedical Cell BioPhysics, University of Twente, Hallenweg 23, 7522 NH Enschede, The NetherlandsVyCAP B.V., Abraham Rademakerstraat 41, 7425 PG Deventer, The NetherlandsDepartment of Medical Oncology, Erasmus MC Cancer Institute and Cancer Genomics Netherlands, Dr. Molewaterplein 40, 3015 GD Rotterdam, the NetherlandsDepartment of Medical Oncology, Erasmus MC Cancer Institute and Cancer Genomics Netherlands, Dr. Molewaterplein 40, 3015 GD Rotterdam, the NetherlandsDepartment of Medical Oncology, Erasmus MC Cancer Institute and Cancer Genomics Netherlands, Dr. Molewaterplein 40, 3015 GD Rotterdam, the NetherlandsMedical Cell BioPhysics, University of Twente, Hallenweg 23, 7522 NH Enschede, The NetherlandsThe 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<sup>®</sup> 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.https://www.mdpi.com/1422-0067/20/3/477CTCself-seeding microwellssingle cell isolationbreast cancerprostate cancer |
| spellingShingle | 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 Self-Seeding Microwells to Isolate and Assess the Viability of Single Circulating Tumor Cells International Journal of Molecular Sciences CTC self-seeding microwells single cell isolation breast cancer prostate cancer |
| title | Self-Seeding Microwells to Isolate and Assess the Viability of Single Circulating Tumor Cells |
| title_full | Self-Seeding Microwells to Isolate and Assess the Viability of Single Circulating Tumor Cells |
| title_fullStr | Self-Seeding Microwells to Isolate and Assess the Viability of Single Circulating Tumor Cells |
| title_full_unstemmed | Self-Seeding Microwells to Isolate and Assess the Viability of Single Circulating Tumor Cells |
| title_short | Self-Seeding Microwells to Isolate and Assess the Viability of Single Circulating Tumor Cells |
| title_sort | self seeding microwells to isolate and assess the viability of single circulating tumor cells |
| topic | CTC self-seeding microwells single cell isolation breast cancer prostate cancer |
| url | https://www.mdpi.com/1422-0067/20/3/477 |
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