Detection and enrichment of disseminated renal carcinoma cells from peripheral blood by immunomagnetic cell separation

We have established an immunomagnetic separation procedure for the detection of circulating tumor cells in the peripheral blood based on the magnetic cell sorting (MACS) technique. In previous in vitro experiments, renal‐cell carcinoma (RCC) cells were mixed with peripheral blood. In dilutions of 1:200 to 1:10 7 tumor cells per mononuclear blood cells, an average recovery rate of 84% of tumor cells was determined. In our study, 104 peripheral blood samples from 59 renal carcinoma patients were analyzed. MACS resulted in significant depletion of leukocytes, permitting a search for tumor cells on just 1 slide. Analyzing 8 ml of peripheral blood per patient, 19/59 RCC patients carried disseminated tumor cells (32%) in the range of 1 to 38 cells (median 8). Interestingly, for the cytokeratin‐positive (CK+) patient group, we found a correlation between tumor cell number and grading (G2 vs. G3) and an increased number of CK+ patients with advanced tumor stage. MACS appears to be an efficient technique to detect disseminated tumor cells in peripheral blood. © 2001 Wiley‐Liss, Inc.

[1]  R Handgretinger,et al.  Detection of circulating melanoma cells by immunomagnetic cell sorting , 1999, Journal of clinical laboratory analysis.

[2]  F. Shanahan,et al.  Micrometastases: marker of metastatic potential or evidence of residual disease? , 1997, Gut.

[3]  M. Ogawa,et al.  DETECTION OF CIRCULATING TUMOR CELLS IN PATIENTS WITH NON – SMALL CELL LUNG CANCER UNDERGOING LOBECTOMY BY VIDEO-ASSISTED THORACIC SURGERY : A POTENTIAL HAZARD FOR INTRAOPERATIVE , 2000 .

[4]  M. Silverman,et al.  Multiple retroperitoneal schwannomas: case report and review of the literature. , 1994, The Journal of urology.

[5]  K Okinaga,et al.  Detection of tumor cells in blood using CD45 magnetic cell separation followed by nested mutant allele‐specific amplification of p53 and K‐ras genes in patients with colorectal cancer , 2000, International journal of cancer.

[6]  R. Cote,et al.  Detection and clinical importance of micrometastatic disease. , 1999, Journal of the National Cancer Institute.

[7]  M. Partridge,et al.  Immunomagnetic separation for enrichment and sensitive detection of disseminated tumour cells in patients with head and neck SCC , 1999, The Journal of pathology.

[8]  Benjamin Geiger,et al.  The catalog of human cytokeratins: Patterns of expression in normal epithelia, tumors and cultured cells , 1982, Cell.

[9]  K. Pantel,et al.  Limitations of reverse-transcriptase polymerase chain reaction analyses for detection of micrometastatic epithelial cancer cells in bone marrow. , 1997, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[10]  M. Rubin,et al.  The detection of renal carcinoma cells in the peripheral blood with an enhanced reverse transcriptase–polymerase chain reaction assay for MN/CA9 , 1999, Cancer.

[11]  P. Hermanek Disseminated tumor cells versus micrometastasis: definitions and problems. , 1999, Anticancer research.

[12]  W. Hrushesky,et al.  Circadian coordination of cancer growth and metastatic spread , 1999, International journal of cancer.

[13]  A. Weiss,et al.  Detection and characterization of carcinoma cells in the blood. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[14]  H. Juhl,et al.  Comparative analysis of bone marrow and venous blood isolates from gastrointestinal cancer patients for the detection of disseminated tumor cells using reverse transcription PCR. , 1997, Cancer research.

[15]  M. C. D. Pinto,et al.  Cytochemical localization of phenol-oxidizing enzymes in lignifying Coleus blumei stems. , 1997 .

[16]  D. Seigneurin,et al.  CK20 gene expression: technical limits for the detection of circulating tumor cells. , 1999, Anticancer research.

[17]  A. Hartmann,et al.  Multiple mutation analyses in single tumor cells with improved whole genome amplification. , 1999, The American journal of pathology.

[18]  K. Pantel,et al.  Phenotypic characteristics of cell lines derived from disseminated cancer cells in bone marrow of patients with solid epithelial tumors: establishment of working models for human micrometastases. , 1999, Cancer research.

[19]  E. Corey,et al.  Detection of disseminated prostate cells by reverse transcription‐polymerase chain reaction (RT‐PCR): Technical and clinical aspects , 1998, International journal of cancer.

[20]  S Miltenyi,et al.  Immunomagnetic enrichment of disseminated epithelial tumor cells from peripheral blood by MACS. , 1998, Experimental hematology.

[21]  H. Juhl,et al.  Disseminated tumor cells in pancreatic cancer patients detected by immunocytology: a new prognostic factor. , 1999, Clinical cancer research : an official journal of the American Association for Cancer Research.

[22]  M. Speicher,et al.  Comparative genomic hybridization, loss of heterozygosity, and DNA sequence analysis of single cells. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[23]  J. Nesland,et al.  Increased sensitivity for detection of micrometastases in bone‐marrow/peripheral‐blood stem‐cell products from breast‐cancer patients by negative immunomagnetic separation , 1998, International journal of cancer.

[24]  A. Dobrovic,et al.  Molecular detection of blood‐borne epithelial cells in colorectal cancer patients and in patients with benign bowel disease , 2000, International journal of cancer.

[25]  K. Schütze,et al.  Isolation by size of epithelial tumor cells : a new method for the immunomorphological and molecular characterization of circulatingtumor cells. , 2000, The American journal of pathology.

[26]  K. Pantel,et al.  Disseminierte Tumorzellen: Diagnostik, prognostische Relevanz, Phänotypisierung und therapeutische Strategien , 1997, Der Chirurg.

[27]  N. Kröger,et al.  Improvement of breast cancer cell detection by immunomagnetic enrichment. , 1999, Cytotherapy.

[28]  D. Sidransky,et al.  Nucleic acid-based methods for the detection of cancer. , 1997, Science.

[29]  J. Minna,et al.  Enrichment of epithelial cells for molecular studies , 1999, Nature Medicine.

[30]  B. Reid,et al.  Loss of heterozygosity analysis using whole genome amplification, cell sorting, and fluorescence-based PCR. , 1999, Genome research.

[31]  A. Dobrovic,et al.  Detection of Circulating Tumor Cells in Colorectal Cancer by Immunobead-PCR Is a Sensitive Prognostic Marker for Relapse of Disease , 1995, Molecular medicine.

[32]  W. Thoenes,et al.  Histopathology and classification of renal cell tumors (adenomas, oncocytomas and carcinomas). The basic cytological and histopathological elements and their use for diagnostics. , 1986, Pathology, research and practice.

[33]  J. Cervós-Navarro,et al.  HEA 125 and Ber EP4: two monoclonal anti-epithelial, non-cytokeratin antibodies distinguishing metastatic carcinomas from glial tumors. , 1993, Clinical neuropathology.

[34]  S. Burchill,et al.  Reliability of reverse transcription-polymerase chain reaction (RT-PCR)-based assays for the detection of circulating tumour cells: a quality-assurance initiative of the EORTC Melanoma Cooperative Group. , 1998, European journal of cancer.

[35]  C. Wittekind,et al.  TNM Klassifikation maligner Tumoren , 1987 .

[36]  T. Schöndorf,et al.  Detection of circulating tumour cells in patients with breast or ovarian cancer by molecular cytogenetics , 1999, British Journal of Cancer.