Isolation of tumor cells using size and deformation
Isolation of tumor cells using size and deformation
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DOI：
10.1016/j.chroma.2009.05.036
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发表时间：
2009-11-20
影响因子：
4.1
通讯作者：
Caggana，Michele
中科院分区：
文献类型：
--
作者：
Mohamed，Hisham;Murray，Megan;Caggana，Michele
从血液中分离和分析循环肿瘤细胞（CTC）是密集研究的主题。虽然可以在分子水平上检测转移，但由于缺乏肿瘤特异性标记物和可预测的DNA异常，进展受到阻碍。这项奋进的主要挑战是可用的感兴趣细胞数量很少，全血中每毫升1-2个。我们已经设计了一种微机械装置，使用物理手段从外周循环中富集和/或分离稀有细胞来破碎全血。它有四个依次变窄的通道阵列，每个通道由一个二维列阵列组成。目前的器件具有宽度为20至5μm、深度为20至5μm的沟道。已经进行了几次优化，导致总共10个衍生设备的制造，只有两种类型用于本研究。两者都具有沿流动轴沿着逐渐变窄的柱之间的间隙宽度，在一个装置上具有20、15、10和5μm的间距。第一个20μm宽的部分分散细胞悬浮液，并在整个装置上产生均匀分布的流动，而其他部分旨在保留越来越小的细胞。整个器件的沟道深度是恒定的，第一种类型是10μm深，第二种类型是20μm深。当将来自八种肿瘤细胞系中的每一种的细胞加载到该装置中时，所有癌细胞都被分离出来。在使用人类全血的混合实验中，我们能够在不受血细胞干扰的情况下破碎癌细胞。此外，可以提取完整的细胞或DNA用于分子分析。这项工作的最终目标是在分子水平上表征细胞，以提供非侵入性方法来监测患者，分期疾病和评估治疗效果。此外，这项工作将使用基因表达谱来深入了解转移。
The isolation and analysis of circulating tumor cells (CTCs) from blood are the subject of intense research. Although tests to detect metastasis on a molecular level are available, progress has been hampered by a lack of tumor-specific markers and predictable DNA abnormalities. The main challenge in this endeavor is the small number of available cells of interest, 1–2 per mL in whole blood. We have designed a micromachined device to fractionate whole blood using physical means to enrich for and/or isolate rare cells from peripheral circulation. It has arrays of four successively narrower channels, each consisting of a two-dimensional array of columns. Current devices have channels ranging in width from 20 to 5μm, and in depth from 20 to 5μm. Several optimizations resulting in the fabrication of a total of 10 derivative devices have been carried out; only two types are used in this study. Both have increasingly narrower gap widths between the columns along the flow axis with 20, 15, 10, and 5μm spacing all on one device. The first 20μm wide segment disperses the cell suspension and creates an evenly distributed flow over the entire device, whereas the others were designed to retain increasingly smaller cells. The channel depth is constant across the entire device, the first type was 10μm deep and the second type is 20μm deep. When cells from each of eight tumor cell lines were loaded into the device, all cancerous cells were isolated. In mixing experiments using human whole blood, we were able to fractionate cancer cells without interference from the blood cells. Additionally, either intact cells, or DNA, could be extracted for molecular analysis. The ultimate goal of this work is to characterize the cells on the molecular level to provide non-invasive methods to monitor patients, stage disease, and assess treatment efficacy. Furthermore, this work will use gene expression profiles to gain insights into metastasis.