Microfluidic antibody arrays for simultaneous cell separation and stimulus.
Microfluidic antibody arrays for simultaneous cell separation and stimulus.
复制标题
用于同时进行细胞分离和刺激的微流控抗体阵列。
DOI：
10.1007/s00216-014-8244-3
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发表时间：
2014
影响因子：
4.3
通讯作者：
Pappas,Dimitri
中科院分区：
文献类型：
--
作者：
Liu,Yan;Germain,Todd;Pappas,Dimitri
开发了包含冲压抗体阵列的微流控芯片，用于同时进行细胞分离和药物测试。使用聚（二甲基硅氧烷）（PDMS）冲压将抗体沉积在微流体通道中，在表面形成离散的细胞捕获区域。然后引入细胞混合物，当使用特异性抗体时导致细胞分离。抗 CD19 抗体区域使 CD19+ Ramos 细胞的捕获纯度达到 94%。捕获多种细胞类型的抗体（例如抗 CD71）也可用于同时捕获多种细胞类型。还可以使用层流将细胞加载到具有空间控制的阵列上。 Ramos B 细胞和 HuT 78 T 细胞均在芯片中分离，并在同一通道中暴露于十字孢菌素。两种细胞系对药物都有相似的反应，根据细胞类型，药物处理 20 小时后，2-10% 的细胞仍然存活。该芯片还可用于分析抗体疗法对癌细胞的功效。抗 CD95 沉积在表面并用于通过外在途径同时捕获细胞和诱导细胞凋亡。与对照抗 CD71 抗体（24 小时后活力为 81%）相比，抗 CD95 表面捕获的细胞活力明显下降（24 小时后活力为 15%）。该芯片可用于各种细胞分离和/或药物测试研究，使研究人员能够分离细胞并针对不同的抗癌化合物测试它们，并使用荧光或其他读出方法跟踪细胞反应。
A microfluidic chip containing stamped antibody arrays was developed for simultaneous cell separation and drug testing. Poly(dimethyl siloxane) (PDMS) stamping was used to deposit antibodies in a microfluidic channel, forming discrete cell-capture regions on the surface. Cell mixtures were then introduced, resulting in the separation of cells when specific antibodies were used. Anti-CD19 antibody regions resulted in 94 % capture purity for CD19+ Ramos cells. An antibody that captures multiple cell types, for example anti-CD71, can also be used to capture several cell types simultaneously. Cells could also be loaded onto the arrays with spatial control using laminar streams. Both Ramos B cells and HuT 78 T cells were isolated in the chip and exposed to staurosporine in the same channel. Both cell lines had similar responses to the drug, with 2–10 % of cells remaining viable after 20 h of drug treatment, depending on cell type. The chip can also be used to analyze the efficacy of antibody therapy against cancer cells. Anti-CD95 was deposited on the surface and used for simultaneous cell capture and apoptosis induction via the extrinsic pathway. Cells captured on anti-CD95 surfaces had significant viability loss (15 % viability after 24 h) when compared with a control anti-CD71 antibody (81 % viability after 24 h). This chip can be used for a variety of cell separation and/or drug testing studies, enabling researchers to isolate cells and test them against different anti-cancer compounds and to follow cell response using fluorescence or other readout methods.