Detection of tumor cells in blood using CD45 magnetic cell separation followed by nested mutant allele-specific amplification ofp53 and K-ras genes in patients with colorectal cancer

Iinuma, Hisae; Okinaga, Kota; Adachi, Miki; Suda, Kazufumi; Sekine, Toshiyuki; Sakagawa, Koichi; Baba, Yasuo; Tamura, Junko; Kumagai, Harumi; Ida, Akimitsu

doi:10.1002/1097-0215(20000720)89:4<337::aid-ijc4>3.0.co;2-r

Cited by 84 publications

(27 citation statements)

References 22 publications

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“…It is possible to widen the cell capture abilities of CTC-antigen–dependent capture methods by using antibody cocktails against cancer type–specific antigens 6,44 . These limitations can be overcome by using methods that enrich aberrant cells from blood by depleting cells that normally exist in blood 13,26,45–52 . This strategy provides tumor antigen–independent enrichment performance, and thus it is applicable to cells disseminated from virtually any tumor type.…”

Section: Introductionmentioning

confidence: 99%

Microfluidic, marker-free isolation of circulating tumor cells from blood samples

et al. 2014

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The ability to isolate and analyze rare circulating tumor cells (CTCs) has the potential to further our understanding of cancer metastasis and enhance the care of cancer patients. In this protocol, we describe the procedure for isolating rare CTCs from blood samples by using tumor antigen–independent microfluidic CTC-iChip technology. The CTC-iChip uses deterministic lateral displacement, inertial focusing and magnetophoresis to sort up to 107 cells/s. By using two-stage magnetophoresis and depletion antibodies against leukocytes, we achieve 3.8-log depletion of white blood cells and a 97% yield of rare cells with a sample processing rate of 8 ml of whole blood/h. The CTC-iChip is compatible with standard cytopathological and RNA-based characterization methods. This protocol describes device production, assembly, blood sample preparation, system setup and the CTC isolation process. Sorting 8 ml of blood sample requires 2 h including setup time, and chip production requires 2–5 d.

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Section: Introductionmentioning

confidence: 99%

Microfluidic, marker-free isolation of circulating tumor cells from blood samples

et al. 2014

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“…11 MACS has been developed to specifically separate rare circulating tumor cells from whole blood for predicting recurrence in patients with solid cancers. 12–14 …”

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confidence: 99%

Magnetic enrichment of bronchial epithelial cells from sputum for lung cancer diagnosis

Qiu

Todd

et al. 2008

Cancer

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BACKGROUND Sputum is an easily accessible diagnostic material for lung cancer early detection by cytologic and molecular genetic analysis of exfoliated airway epithelial cells. However, the use of sputum is limited by its cellular heterogeneity, which includes >95% macrophages and neutrophils and only about 1% bronchial epithelial cells. We propose to obtain concentrated and purified bronchial epithelial cells to improve early detection of lung cancer in sputum samples. METHODS Sputum was collected from patients with stage I nonsmall-cell lung cancer, cancer-free smokers, and healthy nonsmokers. Magnetic-assisted cell sorting (MACS) with anti-CD14 and anti-CD16 antibody beads were used to enrich bronchial epithelial cells by depleting macrophages and neutrophils from sputum. Fluorescence in situ hybridization (FISH) analysis for detection of FHIT deletion and cytology were evaluated in the enriched specimens. RESULTS The bronchial epithelial cells were concentrated to 40% purity from 1.1% of the starting population, yielding an average of 36-fold enrichment and at least 2.3 × 105 cells per sample. Detecting FHIT deletions for lung cancer diagnosis produced 58% sensitivity in the enriched sputum, whereas there was 42% sensitivity in the unenriched samples (P = .02). Cytologic examination of the enriched sputum resulted in 53% sensitivity, as compared with 39% sensitivity in unenriched sputum (P = .03). Furthermore, only 2 cytocentrifuge slides of the unenriched sputum were needed for the analyses, as compared with up to 10 cytocentrifuge slides required from the unprocessed specimens. CONCLUSIONS The enrichment of bronchial epithelial cells could improve the diagnostic value of sputum and the efficiency of genetic and cytologic analysis of lung cancer.

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“…Development of rapid, simple inexpensive and maybe the more importantly specific separation techniques are among the main technological targets in this direction. Magnetic separation has been considered as quite easy, effective and very rapid technique to isolate cells unaltered and viable [12,[15][16][17][18].…”

Section: Resultsmentioning

confidence: 99%

“…Linuma et al [15] have separated CD45 + cells from the cell suspension with commercial MACS microbeads (Miltenyi Biotech, Germany) for the detection of tumor cells in peripheral blood in patients with colorectal cancer. Perez et al [16] have characterized the porcine bone marrow progenitor cells by separating the cells with immunomagnetic techniques.…”

Section: Introductionmentioning

confidence: 99%

Separation of mesenchymal stem cells with magnetic nanosorbents carrying CD105 and CD73 antibodies in flow-through and batch systems

Odabaş

Sayar

Güven

et al. 2008

Journal of Chromatography B

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Detection of tumor cells in blood using CD45 magnetic cell separation followed by nested mutant allele-specific amplification ofp53 and K-ras genes in patients with colorectal cancer

Cited by 84 publications

References 22 publications

Microfluidic, marker-free isolation of circulating tumor cells from blood samples

Microfluidic, marker-free isolation of circulating tumor cells from blood samples

Magnetic enrichment of bronchial epithelial cells from sputum for lung cancer diagnosis

Separation of mesenchymal stem cells with magnetic nanosorbents carrying CD105 and CD73 antibodies in flow-through and batch systems

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