Rare cells

To present theory and methods to determine the effect of sample manipulation on rare cell detection by validating the Poisson theory and measuring cellular abundance by spiking K562 cells with stable EGFP gene expression at known frequencies. After verifying the applicability of this theory, to evaluate the potential impact of red cell lysis on rare cell detection, either using EGFP-K562 or human leukemia cells.
Conclusion:
- Quantification of rare events assessed in terms of cell concentration rather than frequency estimation, may help to achieve robust assays that must be developed and adapted for each patient. - Further experiments will be needed to study the clinical relevance of pathological rare cells in their native sample matrix. - If the malignant rare cell compartment is mainly constituted by ammonium chloride resistant cells, this could lead to an overestimation of CTCs or MRD measurements. - If cell injury derived from sample manipulation can deplete not only healthy, but also pathological cells, including the rare malignant cell compartment, this will also have important implications for the diagnosis and treatment of hematological and nonhematological malignancies. Performance Tracking Beads (daily)