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Current commercially available technology platforms for the enumeration of rare tumor cells in blood or bone marrow are either expensive, inadequately reproducible, or only suited for EPCAM-positive, cytokeratin positive carcinomas. Pathogenesys has elected to enumerate rare tumor cells in the blood or bone marrow of cancer patients after leukocyte-depletion using a proprietary technology that involves the preparation and counting analyte-positive tumor cells on a Cytospin slide. This simplified technique involves leukocyte-depletion of whole, unfractionated blood or marrow using commercially available technology. Reduction of steps leads to higher yields and more reproducible results. Between 90 to 95% of the non-tumor cells are depleted leaving enriched tumor cells at high yield. This technique does not artifically deplete tumor cell clumps, which may be important toward the ultimate assay clinical value. This technique can also be used to enumerate either carcinoma or melanoma cells. The use of a proprietary fixative and lysis buffer after depletion preserves the morphology of the remaining cells, making morphologic analysis of potential tumor cells more definitive.
This approach results in excellent preservation of cellular morphology, as can be seen above. Notice that neutrophils are present and well-preserved. This is due to avoidance of ficol preparations. Competitors’ preparations often contain innumerable smudged or broken cells, making interpretation difficult.
The two above photomicrographs represent bone marrow samples from a patient with breast cancer. Validation Reports will be available on-line soon. For more information, please contact Dr. James Thompson at 949-258-0318 or e-mail at jthompson@pathogenesys.com .
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