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Negative-Selection Enrichment of Circulating Tumor Cells from Peripheral Blood Using the Microfluidic CTC-iChip

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Mammary Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2471))

Abstract

The ability to isolate and analyze rare circulating tumor cells (CTCs) holds the potential to increase our understanding of cancer evolution and allows monitoring of disease and therapeutic responses through a relatively non-invasive blood-based biopsy. While many methods have been described to isolate CTCs from the blood, the vast majority rely on size-based sorting or positive selection of CTCs based on surface markers, which introduces bias into the downstream product by making assumptions about these heterogenous cells. Here we describe a negative-selection protocol for enrichment of CTCs through removal of blood components including red blood cells, platelets, and white blood cells. This procedure results in a product that is amenable to downstream single-cell analytics including RNA-Seq, ATAC-Seq and DNA methylation, droplet digital PCR (ddPCR) for tumor specific transcripts, staining and extensive image analysis, and ex vivo culture of patient-derived CTCs.

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Authors and Affiliations

  1. Massachusetts General Hospital Center for Cancer Research, Harvard Medical School, Charlestown, MA, USA

    Risa Burr, Brian Chirn, Daniel A. Haber & Shyamala Maheswaran

  2. Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

    Jon F. Edd, Avanish Mishra & Mehmet Toner

  3. Howard Hughes Medical Institute, Bethesda, MD, USA

    Daniel A. Haber

  4. Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

    Daniel A. Haber

  5. Shriners Hospital for Children, Boston, MA, USA

    Mehmet Toner

  6. Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

    Mehmet Toner & Shyamala Maheswaran

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  1. Risa Burr
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  2. Jon F. Edd
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  3. Brian Chirn
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  4. Avanish Mishra
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  5. Daniel A. Haber
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  6. Mehmet Toner
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  7. Shyamala Maheswaran
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Correspondence to Shyamala Maheswaran .

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Editors and Affiliations

  1. Cancer Heterogeneity Laboratory, Centre for Cooperative Research in Biosciences (CIC bioGUNE), Basque Research and Technology Alliance (BRTA), Derio, Spain

    Maria dM. Vivanco

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Burr, R. et al. (2022). Negative-Selection Enrichment of Circulating Tumor Cells from Peripheral Blood Using the Microfluidic CTC-iChip. In: Vivanco, M.d. (eds) Mammary Stem Cells. Methods in Molecular Biology, vol 2471. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2193-6_18

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  • DOI: https://doi.org/10.1007/978-1-0716-2193-6_18

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2192-9

  • Online ISBN: 978-1-0716-2193-6

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