Exosome RNA Exosome RNA Research & Industry News
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 researchers from Harvard Medical School 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.
Functional operation of microfluidic CTC isolation requires stabilized whole blood
a Representative images of the micropost array that performs size-based sorting (debulking) in the CTC-iChip. Blood storage in room temperature, even if treated with tiro-EDTA, results in aggregates that contain sheared DNA consistent with cell death and extracellular trap formation. Cold storage without tiro-EDTA leads to clots that contain densely packed platelets (CD61 staining) and intact cells. In both cases, rare cells are trapped within the aggregates. Cold storage with tiro-EDTA consistently permits clean processing (inset). b CTC isolation performance of the CTC-iChip in different storage conditions. All scale bars represent 50 μm. Box-and-whiskers plots show median, interquartile range, maxima, and minima. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 (one-way ANOVA followed by Tukey’s post test)
