Single cell molecular characterization and PD-L1 expression analysis of model CTC using the RareCyte platform
CTCs are exceedingly rare events in blood, which makes a detection platform’s recovery claims hard to verify; this TriCon 2019 poster reports a blinded study in which cells from 8 epithelial lines were spiked into blood as model CTCs and scored on the RareCyte platform for recovery, PD-L1 expression and single-cell mutations.
Presented by RareCyte.
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- A blinded spike-in study across 8 epithelial cell lines recovered 7 of the 8 at frequencies closely correlated with the expected counts. Cultured cells were spiked into blood and carried through the AccuCyte–CyteFinder workflow, with counts reported per 7.5 mL of blood; the single exception was H1573, a highly clumpy line whose recovery was likely skewed by large cell clusters.
- The no-spike controls and the non-epithelial line spikes returned 0 CTCs on unblinding. The melanoma line SK-MEL-28 expresses neither EpCAM nor cytokeratin, so it fell outside the 4-parameter panel used here — a stated scope limit of that panel rather than a detection miss, and the negatives held once the samples were unblinded.
- PD-L1 was measured on the cells themselves, and single-cell sequencing of retrieved cells matched published mutations. Median PD-L1 mean fluorescent intensity spanned more than 20-fold across the panel, from 44 in MCF7 to 979 in SKBR3; individual cells retrieved with the CytePicker module were sequenced with the AmpliSeq Cancer HotSpot Panel on a MiSeq, and the variants called matched published reports for those lines.
