Abstract
Negative enrichment is the preferred approach for tumor cell isolation as it does not rely on biomarker expression. However, size-based negative enrichment methods suffer from well-known recovery/purity trade-off. Non-size based methods have a number of processing steps that lead to compounded cell loss due to extensive sample processing and handling which result in a low recovery efficiency. We present a method that performs negative enrichment in two steps from 2 ml of whole blood in a total assay processing time of 60 min. This negative enrichment method employs upstream immunomagnetic depletion to deplete CD45-positive WBCs followed by a microfabricated filter membrane to perform chemical-free RBC depletion and target cells isolation. Experiments of spiking two cell lines, MCF-7 and NCI-H1975, in the whole blood show an average of >90 % cell recovery over a range of spiked cell numbers. We also successfully recovered circulating tumor cells from 15 cancer patient samples.
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Abbreviations
- CTC:
-
Circulating tumor cells
- WBC:
-
White blood cell
- RBC:
-
Red blood cell
- EMT:
-
Epithelial to mesenchymal transition
- TAC:
-
Tetrameric antibody complexes
- GFP:
-
Green florescent protein
- PBS:
-
Phosphate buffered saline
- EpCAM:
-
Epithelial cell adhesion molecule
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Acknowledgments
This work was supported by the Science and Engineering Research Council of A*STAR (Agency for Science, Technology and Research), Singapore under the grant number 1031490005. In addition, we greatly appreciate the support from Dr. Chaitanya Kantak for microslit membrane fabrication.
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Bhuvanendran Nair Gourikutty Sajay and Chia-Pin Chang contributed equally to this paper.
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Sajay, B.N.G., Chang, CP., Ahmad, H. et al. Microfluidic platform for negative enrichment of circulating tumor cells. Biomed Microdevices 16, 537–548 (2014). https://doi.org/10.1007/s10544-014-9856-2
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DOI: https://doi.org/10.1007/s10544-014-9856-2