Faculty of Engineering and Natural Sciences, Sabanci University, Istanbul, Turkey.
Sabanci University Nanotechnology Research and Application Center, Istanbul, Turkey.
Electrophoresis. 2019 Jan;40(2):315-321. doi: 10.1002/elps.201800324. Epub 2018 Nov 6.
Monocyte heterogeneity and its prevalence are revealed as indicator of several human diseases ranking from cardiovascular diseases to rheumatoid arthritis, chronic kidney diseases, autoimmune multiple sclerosis, and stroke injuries. When monocytes and macrophages are characterized and isolated with preserved genetic, phenotypic and functional properties, they can be used as label-free biomarkers for precise diagnostics and treatment of various diseases. Here, the dielectrophoretic responses of the monocytes and macrophages were examined. We present 3D carbon-electrode dielectrophoresis (carbon-DEP) as a separation tool for U937 monocytes and U937 monocyte-differentiated macrophages. The carbon-electrodes advanced the usability and throughput of DEP separation, presented wider electrochemical stability. Using the 3D carbon-DEP chip, we first identified the selective positive and negative DEP responses and specific crossover frequencies of monocytes and macrophages as their signatures for separation. The crossover frequency of monocytes and macrophages was 17 and 30 kHz, respectively. Next, we separated monocyte and macrophage subpopulations using their specific dielectrophoretic responses. Afterward, we used a fluorescence-activated cell sorter to confirm our results. Finally, we enriched 70% of monocyte cells from the mixed cell population, in other words, concentration of monocyte cells to macrophage cells was five times increased, using the 30-kHz, 10-Vpp electric field and 1 μL/min flow rate.
单核细胞异质性及其流行率被揭示为多种人类疾病的指标，从心血管疾病到类风湿性关节炎、慢性肾病、自身免疫性多发性硬化症和中风损伤。当单核细胞和巨噬细胞以保留遗传、表型和功能特性的方式进行特征化和分离时，它们可以用作各种疾病精确诊断和治疗的无标记生物标志物。在这里，研究了单核细胞和巨噬细胞的介电泳响应。我们提出了 3D 碳电极介电泳（carbon-DEP）作为 U937 单核细胞和 U937 单核细胞分化的巨噬细胞的分离工具。碳电极提高了 DEP 分离的可用性和吞吐量，呈现出更宽的电化学稳定性。使用 3D 碳-DEP 芯片，我们首先确定了单核细胞和巨噬细胞的选择性正和负介电泳响应以及特定的交叉频率作为它们分离的特征。单核细胞和巨噬细胞的交叉频率分别为 17 和 30 kHz。接下来，我们利用它们特定的介电泳响应来分离单核细胞和巨噬细胞亚群。然后，我们使用荧光激活细胞分选器来验证我们的结果。最后，我们使用 30 kHz、10-Vpp 电场和 1 μL/min 的流速从混合细胞群中富集了 70%的单核细胞，换句话说，单核细胞的浓度是巨噬细胞的五倍。
