Comparison of two density gradient centrifugation systems for the enrichment of disseminated tumor cells in blood

Rosenberg, Robert D.; Gertler, Ralf; Friederichs, Jan; Fuehrer, Katrin; Dahm, Michael; Phelps, Richard G.; Thorban, Stefan; Nekarda, H.; Siewert, J. R.

doi:10.1002/cyto.10161

Cited by 257 publications

(212 citation statements)

References 23 publications

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“…Blood samples (25 ml) were fractionated by density-gradient centrifugation at 1,600 ϫ g on OncoQuick columns (Greiner Bio-One, Longwood, FL) (15). The procedure yields three fractions: sedimented blood cells; a low-density fraction in which colorectal cancer (15) and LAM (Table 1) cells were identified; and a fraction above that contains platelets. Cells in urine and chylous fluids were sedimented at 200 ϫ g for 10 min, then washed twice in PBS.…”

Section: Methodsmentioning

confidence: 99%

“…We looked for LAM cells in blood and other body fluids, in particular, those derived from involved organs (urine͞kidney) or vessels (chyle͞ lymphatics). Isolation of cells from blood was performed by using the OncoQuick density gradient system that is designed to prepare samples enriched in circulating cancer cells (15). We report here the isolation of neoplastic LAM cells, identified by TSC2 LOH, from the blood and other body fluids of LAM patients.…”

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confidence: 99%

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Molecular and genetic analysis of disseminated neoplastic cells in lymphangioleiomyomatosis

Crooks

Pacheco–Rodriguez²,

DeCastro³

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

175

122

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Molecular and genetic analysis of disseminated neoplastic cells in lymphangioleiomyomatosis

Crooks

Pacheco–Rodriguez²,

DeCastro³

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

175

122

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“…Inexpensive, easy-to-use (Weitz et al, 1998) OncoQuick Density, Size Porous membrane above separation media for additional separation by filtration (Balic et al, 2005;Clawson et al, 2012;Lagoudianakis et al, 2009;M€ uller et al, 2005;Obermayr et al, 2010;Rosenberg et al, 2002) (Campton et al, 2015) Biophysical e Microfiltration in Two and Three Dimensions Size-based cell separation using pores or three-dimensional geometries.…”

Section: Densitymentioning

confidence: 99%

Circulating tumor cell technologies

2016

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Cancer CTC capture CTC clustersImmunoaffinity enrichment A B S T R A C TCirculating tumor cells, a component of the "liquid biopsy", hold great potential to transform the current landscape of cancer therapy. A key challenge to unlocking the clinical utility of CTCs lies in the ability to detect and isolate these rare cells using methods amenable to downstream characterization and other applications. In this review, we will provide an overview of current technologies used to detect and capture CTCs with brief insights into the workings of individual technologies. We focus on the strategies employed by different platforms and discuss the advantages of each. As our understanding of CTC biology matures, CTC technologies will need to evolve, and we discuss some of the present challenges facing the field in light of recent data encompassing epithelial-to-mesenchymal transition, tumor-initiating cells, and CTC clusters.

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“…Most current methods do not seem to be sensitive or specific enough to detect circulating cells in significant numbers of patients with carcinomas (15)(16)(17). Recently, a major advance in this area occurred with the advent of flow cytometry, which makes possible the detection of CTCs in a good balance of sensitivity and specificity.…”

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confidence: 99%

Detection of circulating tumor cells in breast cancer patients utilizing multiparameter flow cytometry and assessment of the prognosis of patients in different CTCs levels

Fan

Zheng

et al. 2010

Cytometry Pt A

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We wanted to demonstrate the value of multiparameter flow cytometry in detecting human tumor cells of breast cancer (BC) (SKBR-3) in normal peripheral blood. In addition, we investigated a cluster of patients to compare the overall survival (OS) between advanced BC patients [circulating tumor cells (CTCs) !5 group] and limited BC patients (CTCs <5 group). SKBR-3 human BC cells were serially diluted in normal whole blood to demonstrate the sensitivity of multiparameter flow cytometry for detecting CTCs, and we also compared the specificity with reverse transcriptase polymerase chain reaction (RT-PCR) method. On the other hand, we detected CTCs among 45 patients by multiparameter flow cytometry. OS was calculated by the Kaplan-Meier product limit method, and compared it between CTCs <5 and CTCs !5 groups with the log-rank test. Cox regression models were fitted to determine the associated factors on survival. Human BC cells (SKBR-3) could be differentiated from normal blood based on the multiple light scatter and cell surface marker expression by multiparameter flow cytometry. The method was found to have a sensitivity limit of 10 25 and was effective for detecting human BC cells in vivo. It also found that this method had a higher specificity compared with RT-PCR. For the retrospective study, the median OS was 95 weeks and 65.5 weeks (P < 0.05, 2-tailed) for patients with CTCs <5 and CTCs !5, respectively. Kaplan-Meier was used to analyze the patients' survival with Log Rank P = 0.004 and Breslow P = 0.003, which showed that these two groups had statistically significant difference. Cox regression analysis was performed, and we found CTCs levels, metastasis and age (P < 0.05) were three relative factors for patients' survival. Multiparameter flow cytometry can detect CTCs effectively and has the potential to be a valuable tool for prognosis assessment among BC patients in clinical situations in China. '

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Comparison of two density gradient centrifugation systems for the enrichment of disseminated tumor cells in blood

Abstract: OncoQuick significantly reduced the co-enriched number of MNCs, with a high tumor cell recovery rate. Processing blood from tumor patients with OncoQuick increased the chance of detecting circulating tumor cells.

Cited by 257 publications

References 23 publications

Molecular and genetic analysis of disseminated neoplastic cells in lymphangioleiomyomatosis

Molecular and genetic analysis of disseminated neoplastic cells in lymphangioleiomyomatosis

Circulating tumor cell technologies

Detection of circulating tumor cells in breast cancer patients utilizing multiparameter flow cytometry and assessment of the prognosis of patients in different CTCs levels

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