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Nat Commun. 2017 Nov 23;8(1):1733. doi: 10.1038/s41467-017-01705-y.

Whole blood stabilization for the microfluidic isolation and molecular characterization of circulating tumor cells.

Author information

1
BioMEMS Resource Center, Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA.
2
Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA.
3
Cancer Center, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA.
4
Department of Radiation Oncology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA.
5
Department of Pathology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA.
6
Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA.
7
Howard Hughes Medical Institute, Chevy Chase, MD, 20815, USA.
8
BioMEMS Resource Center, Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA. sstott@mgh.harvard.edu.
9
Cancer Center, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA. sstott@mgh.harvard.edu.
10
Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA. sstott@mgh.harvard.edu.
11
BioMEMS Resource Center, Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA. mtoner@hms.harvard.edu.
12
Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA. mtoner@hms.harvard.edu.

Abstract

Precise rare-cell technologies require the blood to be processed immediately or be stabilized with fixatives. Such restrictions limit the translation of circulating tumor cell (CTC)-based liquid biopsy assays that provide accurate molecular data in guiding clinical decisions. Here we describe a method to preserve whole blood in its minimally altered state by combining hypothermic preservation with targeted strategies that counter cooling-induced platelet activation. Using this method, whole blood preserved for up to 72 h can be readily processed for microfluidic sorting without compromising CTC yield and viability. The tumor cells retain high-quality intact RNA suitable for single-cell RT-qPCR as well as RNA-Seq, enabling the reliable detection of cancer-specific transcripts including the androgen-receptor splice variant 7 in a cohort of prostate cancer patients with an overall concordance of 92% between fresh and preserved blood. This work will serve as a springboard for the dissemination of diverse blood-based diagnostics.

PMID:
29170510
PMCID:
PMC5700979
DOI:
10.1038/s41467-017-01705-y
[Indexed for MEDLINE]
Free PMC Article

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