Clinically Relevant Microfluidic Magnetophoretic Isolation of Rare-Cell Populations for Diagnostic and Therapeutic Monitoring Applications
Clinically Relevant Microfluidic Magnetophoretic Isolation of Rare-Cell Populations for Diagnostic and Therapeutic Monitoring Applications
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DOI：
10.1021/ac2022844
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发表时间：
2012-02-07
影响因子：
7.4
通讯作者：
Murthy, Shashi K.
中科院分区：
文献类型：
--
作者：
Plouffe, Brian D.;Mahalanabis, Madhumita;Murthy, Shashi K.
具有生物医学意义的细胞，尽管其功能重要性，但通常以非常小的数量存在。因此，分析和分离以前难以获得的稀有细胞，如外周造血干细胞、内皮祖细胞或循环肿瘤细胞，需要不损害细胞活力的有效、灵敏和特异的程序。目前的研究建立在以前的工作上的合理设计的微流控磁泳细胞分离平台能够240 μ L min（-1）的吞吐量。首先使用MCF-7（1-1000个总细胞）作为靶稀有细胞加标到高浓度Raji B淋巴细胞非靶细胞（类似于10（6）个总细胞）中进行概念验证。这些实验建立了基于磁性的分离方法，可直接从全血中分离50个MCF-7细胞。结果表明，收集效率大于85%，纯度大于90%。接下来，以快速和有效的方式（>96%）直接从人全血中分离驻留的内皮祖细胞和造血干细胞。两个细胞群可以同时分离，并通过免疫荧光染色，单独识别和计数。总的来说，所提出的装置说明了一种可行的分离平台，用于直接从全血样品中高纯度、高效和快速地收集稀有细胞群。
Cells of biomedical interest are, despite their functional significance, often present in very small numbers. Therefore the analysis and isolation of previously inaccessible rare cells, such as peripheral hematopoietic stem cells, endothelial progenitor cells, or circulating tumor cells, require efficient, sensitive, and specific procedures that do not compromise the viability of the cells. The current study builds on previous work on a rationally designed microfluidic magnetophoretic cell separation platform capable of throughputs of 240 mu L min(-1). Proof-of-concept was first conducted using MCF-7 (1-1000 total cells) as the target rare cell spiked into high concentrations of Raji B-lymphocyte nontarget cells (similar to 10(6) total cells). These experiments lead to the establishment of a magnet-based separation for the isolation of 50 MCF-7 cells directly from whole blood. Results show an efficiency of collection greater than 85%, with a purity of over 90%. Next, resident endothelial progenitor cells and hematopoietic stem cells are directly isolated from whole human blood in a rapid and efficient fashion (>96%). Both cell populations could be simultaneously isolated and, via immunofluorescent staining, individually identified and enumerated. Overall, the presented device illustrates a viable separation platform for high purity, efficient, and rapid collection of rare cell populations directly from whole blood samples.