Microfluidic inertia enhanced phase partitioning for enriching nucleated cell populations in blood
Microfluidic inertia enhanced phase partitioning for enriching nucleated cell populations in blood
复制标题
微流体惯性增强相分配以富集血液中的有核细胞群
DOI：
10.1039/c2lc40663b
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发表时间：
2013
期刊：
影响因子：
6.1
通讯作者：
Sethu, Palaniappan
中科院分区：
文献类型：
--
作者：
Parichehreh, Vahidreza;Medepallai, Krishnakiran;Babbarwal, Karan;Sethu, Palaniappan
血液中的有核细胞，如白细胞（WBC）和其他稀有细胞，包括外周血干细胞（PBSC）和循环肿瘤细胞（CTC），对于患者监测和临床诊断具有重要价值。在不使用抗体或离心的情况下，使用无标记技术从污染的红细胞 (RBC) 中富集有核细胞是非常理想的，以确保最大限度地减少细胞损失和激活。为了实现这一目标，我们展示了一种新的微流体技术的概念验证，该技术将水相分配与惯性聚焦相结合，以实现血液中有核细胞的富集。该技术利用红细胞与葡聚糖相 (DEX) 的选择性亲和力来实现初始分离，这种分离通过高纵横比通道中产生的惯性力而放大。在我们的实验中，我们以 500 : 1（RBC : 有核细胞）的比例加入代表性有核细胞、MOLT-3 细胞（人、外周血、T 淋巴母细胞系）和 MCF-7 细胞（人乳腺癌细胞系），并完成了约 96% 的红细胞的去除，同时保留了约 98% 的有核细胞。通过使用相同的过程对富集的有核细胞混合物进行第二次处理，可以实现更高的纯度。第二遍进一步增强了红细胞的去除（> 初始浓度的 99%），而有核细胞则被回收而没有任何进一步的损失。因此，该技术有可能单独使用或作为临床和研究环境中的样品制备工具用于各种临床和研究应用。
Nucleated cells in blood like white blood cells (WBCs) and other rare cells including peripheral blood stem cells (PBSCs) and circulating tumor cells (CTCs) possess significant value for patient monitoring and clinical diagnosis. Enrichment of nucleated cells from contaminating red blood cells (RBCs) using label-free techniques without the use of antibodies or centrifugation is highly desirable to ensure minimal cell loss and activation. To accomplish this, we demonstrate proof-of-concept of a new microfluidic technique that combines aqueous phase partitioning with inertial focusing to accomplish enrichment of nucleated cells in blood. This technique exploits selective affinity of RBCs to the dextran phase (DEX) to accomplish initial separation which is amplified by inertial forces that develop in high-aspect-ratio channels. In our experiments, we spiked RBC samples with representative nucleated cells, MOLT-3 cells (human, peripheral blood, T lymphoblast cell line) and MCF-7 cells (human breast cancer cell line) in a ratio of 500 : 1 (RBCs : nucleated cells) and accomplished depletion of ∼96% of RBCs while retaining ∼98% of nucleated cells. Higher purity can be accomplished by subjecting the enriched nucleated cell mixture to a second pass via the same process. The second pass further enhances RBC depletion (>99% of initial concentration) whereas nucleated cells were recovered without any further loss. This technique therefore has the potential to be utilized either alone or as a sample preparation tool in the clinical and research setting for various clinical and research applications.