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

Microfluidic, marker-free isolation of circulating tumor cells from blood samples.
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DOI:
10.1038/nprot.2014.044
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发表时间:
2014-03
期刊:
影响因子:
14.8
通讯作者:
--
中科院分区:
生物学1区
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--
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分离和分析罕见的循环肿瘤细胞(CTCs)的能力有可能增进我们对癌症转移的理解,并改善癌症患者的治疗。在本实验方案中,我们描述了利用不依赖肿瘤抗原的微流控CTC - iChip技术从血液样本中分离罕见CTCs的步骤。CTC - iChip利用确定性侧向位移、惯性聚焦和磁泳技术,每秒可分选多达10⁷个细胞。通过使用两级磁泳以及抗白细胞的去除抗体,我们实现了白细胞3.8个对数级的去除,罕见细胞的回收率达到97%,全血样本处理速率为每小时8毫升。CTC - iChip与标准的细胞病理学和基于RNA的鉴定方法兼容。本实验方案描述了设备的生产、组装、血液样本的制备、系统设置以及CTC分离过程。分选8毫升血液样本(包括设置时间)需要2小时,芯片生产需要2 - 5天。
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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