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

Author

Listed:
  • Keith H. K. Wong

    (Harvard Medical School
    Harvard Medical School)

  • Shannon N. Tessier

    (Harvard Medical School
    Harvard Medical School)

  • David T. Miyamoto

    (Harvard Medical School
    Harvard Medical School)

  • Kathleen L. Miller

    (Harvard Medical School
    Harvard Medical School)

  • Lauren D. Bookstaver

    (Harvard Medical School
    Harvard Medical School)

  • Thomas R. Carey

    (Harvard Medical School
    Harvard Medical School)

  • Cleo J. Stannard

    (Harvard Medical School
    Harvard Medical School)

  • Vishal Thapar

    (Harvard Medical School
    Harvard Medical School)

  • Eric C. Tai

    (Harvard Medical School)

  • Kevin D. Vo

    (Harvard Medical School)

  • Erin S. Emmons

    (Harvard Medical School)

  • Haley M. Pleskow

    (Harvard Medical School)

  • Rebecca D. Sandlin

    (Harvard Medical School
    Harvard Medical School)

  • Lecia V. Sequist

    (Harvard Medical School
    Harvard Medical School)

  • David T. Ting

    (Harvard Medical School
    Harvard Medical School)

  • Daniel A. Haber

    (Harvard Medical School
    Harvard Medical School
    Howard Hughes Medical Institute)

  • Shyamala Maheswaran

    (Harvard Medical School
    Harvard Medical School)

  • Shannon L. Stott

    (Harvard Medical School
    Harvard Medical School
    Harvard Medical School)

  • Mehmet Toner

    (Harvard Medical School
    Harvard Medical School)

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.

Suggested Citation

  • Keith H. K. Wong & Shannon N. Tessier & David T. Miyamoto & Kathleen L. Miller & Lauren D. Bookstaver & Thomas R. Carey & Cleo J. Stannard & Vishal Thapar & Eric C. Tai & Kevin D. Vo & Erin S. Emmons , 2017. "Whole blood stabilization for the microfluidic isolation and molecular characterization of circulating tumor cells," Nature Communications, Nature, vol. 8(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_s41467-017-01705-y
    DOI: 10.1038/s41467-017-01705-y
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