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Detection of EpCAM-Negative and Cytokeratin-Negative Circulating Tumor Cells in Peripheral Blood.

Mikolajczyk SD, Millar LS, Tsinberg P, Coutts SM, Zomorrodi M, Pham T, Bischoff FZ, Pircher TJ - J Oncol (2011)

Bottom Line: Higher recovery of CTCs was demonstrated using antibody mixtures compared to anti-EpCAM.In addition, CK-positive breast cancer cells were found in 15 of 24 samples (63%; range 1-60 CTCs), while all samples contained additional CE-positive cells (range 1-41; median = 11; P = .02).Thus, antibody mixtures against a range of cell surface antigens enables capture of more CTCs than anti-EpCAM alone and CE staining enables the detection of CK-negative CTCs.

View Article: PubMed Central - PubMed

Affiliation: Research and Development, Biocept Inc., 5810 Nancy Ridge Drive, Suite 150, San Diego, CA 92121, USA.

ABSTRACT
Enrichment of rare circulating tumor cells (CTCs) in blood is typically achieved using antibodies to epithelial cell adhesion molecule (EpCAM), with detection using cytokeratin (CK) antibodies. However, EpCAM and CK are not expressed in some tumors and can be downregulated during epithelial-to-mesenchymal transition. A micro-fluidic system, not limited to EpCAM or CK, was developed to use multiple antibodies for capture followed by detection using CEE-Enhanced (CE), a novel in situ staining method that fluorescently labels the capture antibodies bound to CTCs. Higher recovery of CTCs was demonstrated using antibody mixtures compared to anti-EpCAM. In addition, CK-positive breast cancer cells were found in 15 of 24 samples (63%; range 1-60 CTCs), while all samples contained additional CE-positive cells (range 1-41; median = 11; P = .02). Thus, antibody mixtures against a range of cell surface antigens enables capture of more CTCs than anti-EpCAM alone and CE staining enables the detection of CK-negative CTCs.

No MeSH data available.


Related in: MedlinePlus

Diagram of the CEE microchannel. (a) Top view of the channel showing the inlet where sample is loaded and the outlet that is attached to a syringe pump to draw sample through the channel. (b) Bottom view shows the area where 9,000 posts are located in the silicone block and the channel sealed with the bottom cover slip. The total volume of the microchannel is 24 μL. A standard microscope slide is added for stability during handling but is removed to visualize cells. The microchannel is inverted on a microscope and the captured cells viewed through the coverslip.
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Related In: Results  -  Collection


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fig1: Diagram of the CEE microchannel. (a) Top view of the channel showing the inlet where sample is loaded and the outlet that is attached to a syringe pump to draw sample through the channel. (b) Bottom view shows the area where 9,000 posts are located in the silicone block and the channel sealed with the bottom cover slip. The total volume of the microchannel is 24 μL. A standard microscope slide is added for stability during handling but is removed to visualize cells. The microchannel is inverted on a microscope and the captured cells viewed through the coverslip.

Mentions: CEE microchannel design is illustrated in Figure 1. The random size and spacing of the posts is mathematically designed to avoid laminar flow through the channel, thus maximizing cell contact with the inner surfaces. The entire inner surface of the channel is derivatized with tethered streptavidin and therefore cells may be specifically bound on any surface. In practice the majority of the CTCs are captured on the posts though some cells are found on the channel floor.


Detection of EpCAM-Negative and Cytokeratin-Negative Circulating Tumor Cells in Peripheral Blood.

Mikolajczyk SD, Millar LS, Tsinberg P, Coutts SM, Zomorrodi M, Pham T, Bischoff FZ, Pircher TJ - J Oncol (2011)

Diagram of the CEE microchannel. (a) Top view of the channel showing the inlet where sample is loaded and the outlet that is attached to a syringe pump to draw sample through the channel. (b) Bottom view shows the area where 9,000 posts are located in the silicone block and the channel sealed with the bottom cover slip. The total volume of the microchannel is 24 μL. A standard microscope slide is added for stability during handling but is removed to visualize cells. The microchannel is inverted on a microscope and the captured cells viewed through the coverslip.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3090615&req=5

fig1: Diagram of the CEE microchannel. (a) Top view of the channel showing the inlet where sample is loaded and the outlet that is attached to a syringe pump to draw sample through the channel. (b) Bottom view shows the area where 9,000 posts are located in the silicone block and the channel sealed with the bottom cover slip. The total volume of the microchannel is 24 μL. A standard microscope slide is added for stability during handling but is removed to visualize cells. The microchannel is inverted on a microscope and the captured cells viewed through the coverslip.
Mentions: CEE microchannel design is illustrated in Figure 1. The random size and spacing of the posts is mathematically designed to avoid laminar flow through the channel, thus maximizing cell contact with the inner surfaces. The entire inner surface of the channel is derivatized with tethered streptavidin and therefore cells may be specifically bound on any surface. In practice the majority of the CTCs are captured on the posts though some cells are found on the channel floor.

Bottom Line: Higher recovery of CTCs was demonstrated using antibody mixtures compared to anti-EpCAM.In addition, CK-positive breast cancer cells were found in 15 of 24 samples (63%; range 1-60 CTCs), while all samples contained additional CE-positive cells (range 1-41; median = 11; P = .02).Thus, antibody mixtures against a range of cell surface antigens enables capture of more CTCs than anti-EpCAM alone and CE staining enables the detection of CK-negative CTCs.

View Article: PubMed Central - PubMed

Affiliation: Research and Development, Biocept Inc., 5810 Nancy Ridge Drive, Suite 150, San Diego, CA 92121, USA.

ABSTRACT
Enrichment of rare circulating tumor cells (CTCs) in blood is typically achieved using antibodies to epithelial cell adhesion molecule (EpCAM), with detection using cytokeratin (CK) antibodies. However, EpCAM and CK are not expressed in some tumors and can be downregulated during epithelial-to-mesenchymal transition. A micro-fluidic system, not limited to EpCAM or CK, was developed to use multiple antibodies for capture followed by detection using CEE-Enhanced (CE), a novel in situ staining method that fluorescently labels the capture antibodies bound to CTCs. Higher recovery of CTCs was demonstrated using antibody mixtures compared to anti-EpCAM. In addition, CK-positive breast cancer cells were found in 15 of 24 samples (63%; range 1-60 CTCs), while all samples contained additional CE-positive cells (range 1-41; median = 11; P = .02). Thus, antibody mixtures against a range of cell surface antigens enables capture of more CTCs than anti-EpCAM alone and CE staining enables the detection of CK-negative CTCs.

No MeSH data available.


Related in: MedlinePlus