Liquid Biopsy Using The Nanotube-CTC- Chip
PublicIn this work, we describe the development of the nanotube-CTC-chip for capture and enumeration of circulating tumor cells (CTC) in breast cancer patients. The nanotube-CTC-chip is a new 76-element microarray technology that combines carbon nanotube surfaces with microarray batch manufacturing techniques for capture and isolation of tumor-derived epithelial cells.\n\nUsing a combination of red blood cell (RBC) lysis and preferential adherence, we demonstrate the capture and enrichment of CTCs with 5-log reduction of contaminating WBCs. EpCAM-MDA-MB-231/Luciferase-2A-Green Fluorescent Protein (GFP) cells were spiked in the blood of wild mice and enriched using an RBC lysis protocol. The enriched samples were then processed using the nanotube-CTC-chip for preferential CTC adherence on the nanosurface and counting the GFP cells enabled anywhere from 89-100% capture from the droplets.\n\nElectron microscopy (EM) studies revealed strong focal adhesion with filaments from the cell body to the nanotube surface. The preferential adherence strategy of CTC capture compared to collagen adhesion matrix (CAM) scaffolding method, which in the past reported as a viable strategy for CTC capture in patients. The CAM scaffolding on the device surface yielded 50% adherence with 100% tracking of cancer cells (adhered and non-adhered) versus carbon nanotubes with >90% adherence and 100% tracking for the same protocol.\n\nThe nanotube-CTC-chip successfully captured CTCs in the peripheral blood of breast cancer patients (Stage 1-4) with a range of 4–238 CTCs per 8.5 ml blood or 0.5-28 CTCs per ml. CTCs (based on CK8/18, Her2, EGFR) successfully identified in 7/7 breast cancer patients, and no CTCs captured in healthy controls (n=2). CTC enumeration based on different markers using the nanotube-CTC-chip enables dynamic views of metastatic progression.
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- etd-052719-201415
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- 2019
- Date created
- 2019-05-27
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- Rights statement
- Dernière modification
- 2021-01-05
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