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

Wong, K.H.K., Tessier, S.N., Miyamoto, D.T. et al.

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.

Citation

Wong, K.H.K., Tessier, S.N., Miyamoto, D.T. et al. "Whole blood stabilization for the microfluidic isolation and molecular characterization of circulating tumor cells" Nature Communications (2017): 1,733