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Detection of Circulating Tumor Cells in Blood Using an Optimized Density Gradient Centrifugation

机译:使用优化密度梯度离心检测血液中循环肿瘤细胞

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The aim of the study was to compare the new density gradient centrifugation system OncoQuick with the standard density gradient centrifugation system Ficoll for improved tumor cell enrichment in blood of tumor patients. Evaluation of OncoQuick and Ficoll density gradient centrifugation was performed by flow-cytometry and immunocytochemistry using 10 ml unspiked and tumor cell-spiked blood samples of tumor-free probands. From 10 ml blood, OncoQuick density gradient centrifugation separated a cell fraction which consisted of a mean cell number of 9.5X10~4 mononuclear cells compared to 1.8xl07 cells by Ficoll. Density gradient centrifugation of tumor cell-spiked blood samples with OncoQuick and Ficoll led to similar tumor cell recovery rates, between 70% and 90% for both methods. The improved depletion of mononuclear blood cells by OncoQuick simplified further immunocytochem-ical evaluation of the enriched cell fraction, which could be spun onto 1-2 glass slides by cytocentrifugation. In comparison, the mononuclear cells separated by Ficoll had to be spun onto more than 50 glass slides for complete im-munocytochemical evaluation. Consequently, tumor cell density on each cy-tospin was higher after OncoQuick preparation compared to Ficoll. Density gradient centrifugation with OncoQuick results in higher relative tumor cell enrichment than Ficoll density gradient centrifugation. This simplifies further immunocytochemical tumor cell detection and is a promising tool for the detection of circulating tumor cells in blood of tumor patients.
机译:该研究的目的是将新的密度梯度离心系统进行比较,具有标准密度梯度离心系统Ficoll以改善肿瘤患者血液中的肿瘤细胞富集。通过流式细胞术和免疫细胞化学使用10mL未加速的肿瘤胰腺的肿瘤的血液样品进行砧率和Ficoll密度梯度离心的评估。从10ml血液中,oncoquick密度梯度离心分离,细胞级分,其由Ficoll的1.8×10 7细胞相比,由9.5×10〜4个单核细胞的平均细胞数组成。肿瘤细胞掺入血液样品的密度梯度离心用oncoquick和Ficoll导致相似的肿瘤细胞回收率,两种方法的70%和90%。通过oncoquick改善单核血细胞的耗竭,简化了富集细胞级分的进一步免疫细胞 - ICE评估,这可以通过细胞加压旋转到1-2玻璃载玻片上。相比之下,由Ficoll分离的单核细胞必须旋转到50多个玻璃载玻片上,用于完全IM-MunococoCrochemical评估。因此,与Ficoll相比,在砧率准备后,每个Cy-Tophin的肿瘤细胞密度较高。密度梯度与oncoquick离心,导致比Ficoll密度梯度离心更高的相对肿瘤细胞富集。这简化了进一步的免疫细胞化学肿瘤细胞检测,并且是检测肿瘤患者血液中循环肿瘤细胞的有希望的工具。

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