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

Risa Burr; Jon F Edd; Brian Chirn; Avanish Mishra; Daniel A Haber; Mehmet Toner; Shyamala Maheswaran

doi:10.1007/978-1-0716-2193-6_18

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

Methods Mol Biol. 2022:2471:309-321. doi: 10.1007/978-1-0716-2193-6_18.

Authors

Risa Burr¹, Jon F Edd², Brian Chirn¹, Avanish Mishra², Daniel A Haber^{1

3

4}, Mehmet Toner^{2

5

6}, Shyamala Maheswaran^{7

8}

Affiliations

¹ Massachusetts General Hospital Center for Cancer Research, Harvard Medical School, Charlestown, MA, USA.
² Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
³ Howard Hughes Medical Institute, Bethesda, MD, USA.
⁴ Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
⁵ Shriners Hospital for Children, Boston, MA, USA.
⁶ Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
⁷ Massachusetts General Hospital Center for Cancer Research, Harvard Medical School, Charlestown, MA, USA. Maheswaran@helix.mgh.harvard.edu.
⁸ Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. Maheswaran@helix.mgh.harvard.edu.

PMID: 35175606
DOI: 10.1007/978-1-0716-2193-6_18

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.

Keywords: Blood-based biopsy; Circulating tumor cell; Metastasis; Microfluidics; Negative selection; iChip.

MeSH terms

Cell Count
Cell Line, Tumor
Cell Separation / methods
Humans
Microfluidics / methods
Neoplastic Cells, Circulating* / pathology