使用含有保存试剂的采集装置稳定血液中的循环肿瘤细胞。
Stabilization of circulating tumor cells in blood using a collection device with a preservative reagent.
机构信息
R&D Division, Streck, Inc,, 7002 S 109 Street, La Vista, NE 68128, USA.
出版信息
Cancer Cell Int. 2014 Mar 7;14(1):23. doi: 10.1186/1475-2867-14-23.
BACKGROUND
The enumeration and characterization of circulating tumor cells (CTCs) in the blood of cancer patients is useful for cancer prognostic and treatment monitoring purposes. The number of CTCs present in patient blood is very low; thus, robust technologies have been developed to enumerate and characterize CTCs in patient blood samples. One of the challenges to the clinical utility of CTCs is their inherent fragility, which makes these cells very unstable during transportation and storage of blood samples. In this study we investigated Cell-Free DNA BCT™ (BCT), a blood collection device, which stabilizes blood cells in a blood sample at room temperature (RT) for its ability to stabilize CTCs at RT for an extended period of time.
METHODS
Blood was drawn from each donor into K3EDTA tube, CellSave tube and BCT. Samples were then spiked with breast cancer cells (MCF-7), transported and stored at RT. Spiked cancer cells were counted using the Veridex CellSearch™ system on days 1 and 4. The effect of storage on the stability of proteins and nucleic acids in the spiked cells isolated from K3EDTA tube and BCT was determined using fluorescence staining and confocal laser scanning microscopy.
RESULTS
MCF-7 cell recovery significantly dropped when transported and stored in K3EDTA tubes. However, in blood collected into CellSave tubes and BCTs, the MCF-7 cell count was stable up to 4 days at RT. Epithelial cell adhesion molecule (EpCAM) and cytokeratin (CK) in MCF-7 cells isolated from BCTs was stable at RT for up to 4 days, whereas in MCF-7 cells isolated from K3EDTA blood showed reduced EpCAM and CK protein expression. Similarly, BCTs stabilized c-fos and cyclin D1 mRNAs as compared to K3EDTA tubes.
CONCLUSION
Cell-Free DNA™ BCT blood collection device preserves and stabilizes CTCs in blood samples for at least 4 days at RT. This technology may facilitate the development of new non-invasive diagnostic and prognostic methodologies for CTC enumeration as well as characterization.
背景
在癌症患者的血液中对循环肿瘤细胞（CTC）进行计数和特征分析有助于癌症预后和治疗监测。患者血液中 CTC 的数量非常低；因此，已经开发了强大的技术来对患者血液样本中的 CTC 进行计数和特征分析。CTC 临床应用的一个挑战是其内在的脆弱性，这使得这些细胞在血液样本的运输和储存过程中非常不稳定。在这项研究中，我们研究了 Cell-Free DNA BCT（BCT），这是一种血液采集装置，它可以在室温（RT）下稳定血液样本中的血细胞，从而能够在较长时间内稳定 RT 下的 CTC。
方法
每位供体的血液均采集至 K3EDTA 管、CellSave 管和 BCT 中。然后，将乳腺癌细胞（MCF-7）加入到样本中，在 RT 下进行运输和储存。在第 1 天和第 4 天，使用 Veridex CellSearch™系统对加入的癌细胞进行计数。使用荧光染色和共聚焦激光扫描显微镜确定从 K3EDTA 管和 BCT 中分离出的加入细胞中蛋白质和核酸的储存稳定性。
结果
当 MCF-7 细胞在 K3EDTA 管中运输和储存时，细胞回收量显著下降。然而，在采集到 CellSave 管和 BCT 中的血液中，MCF-7 细胞计数在 RT 下可稳定长达 4 天。从 BCT 中分离的 MCF-7 细胞中的上皮细胞黏附分子（EpCAM）和细胞角蛋白（CK）在 RT 下稳定长达 4 天，而从 K3EDTA 血液中分离的 MCF-7 细胞中 EpCAM 和 CK 蛋白表达减少。同样，BCT 稳定了 c-fos 和细胞周期蛋白 D1mRNA，而 K3EDTA 管则没有。
结论
Cell-Free DNA™ BCT 血液采集装置可在 RT 下至少稳定血液样本中的 CTC 4 天。这项技术可能促进开发新的非侵入性诊断和预后方法，用于 CTC 计数和特征分析。
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