Immunomagnetic enrichment and detection of isolated tumor cells in bone marrow of patients with epithelial malignancies |
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Authors: | Weihrauch Martin R Skibowski Edmund Draube Andreas Geller Andrea Tesch Hans Diehl Volker Bohlen Heribert |
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Institution: | (1) Department of Internal Medicine I of the University of Cologne, Germany;(2) Hematooncological Outpatient Clinic, Frankfurt, Germany;(3) Axiogenesis GmbH Cologne, Germany |
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Abstract: | The detection of isolated tumor cells (ITC) in the bone marrow of patients with epithelial malignancies is an independant
prognostic factor for several entities as breast cancer, colorectal cancer or non-small lung cancer. However, with conventional
immunocytology using Ficoll density gradient and APAAP staining, only a small proportion of the bone marrow samples can be
scanned for cytokeratin-positive (CK+) cells. To improve detection rates, we evaluated the enrichment of ITC by magnetic activated
cell sorting (MACS) compared to regularly stained cytospins. Recovery experiments with a CK+ breast cancer cell line (SKBR3)
were performed to calculate the MACS enrichment rate. Bone marrow was obtained by aspiration from 20 patients with carcinomas
of epithelial origin and from 17 controls. ITC were enriched and stained with magnetically labeled CAM 5.2 antibodies directed
to cytokeratin 7 and 8. MACS of SKBR3 seeded in peripheral blood revealed average recovery rates of 62% and 48% and average
enrichment factors of 104-fold and 8139-fold of the CK+ cells after one and after two separations, respectively. After immunomagnetic
enrichment, CK+ cells were detected in 16 of 20 (80%) cancer patients, whereas only 7 (35%) patients showed CK+ cells without
magnetic enrichment (P=0.002). Ten of twelve (83%) patients with metastatic disease (stage M1) and six of eight (75%) patients without any overt metastases (M0) had CK+ cells in their bone marrow. None of the negative controls showed any CK+ cells. Enrichment with magnetically labeled
anti cytokeratin antibodies increases the detection rate of epithelial cells in bone marrow of cancer patients compared to
immunocytology.
This revised version was published online in July 2006 with corrections to the Cover Date. |
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Keywords: | immunomagnetic isolated tumor cells MACS micrometastasis |
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