Clinically Relevant Microfluidic Magnetophoretic Isolation of Rare-Cell Populations for Diagnostic and Therapeutic Monitoring Applications

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• 作者：Brian D. Plouffe ; Madhumita Mahalanabis ; Laura H. Lewis ; Catherine M. Klapperich ; Shashi K. Murthy
• 刊名：Analytical Chemistry
• 出版年：2012
• 出版时间：February 7, 2012
• 年：2012
• 卷：84
• 期：3
• 页码：1336-1344
• 全文大小：340K
• 年卷期：v.84,no.3(February 7, 2012)
• ISSN：1520-6882

文摘

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 渭L min^鈥?. Proof-of-concept was first conducted using MCF-7 (1鈥?000 total cells) as the target rare cell spiked into high concentrations of Raji B-lymphocyte nontarget cells (10⁶ 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.