Okşak Nilgün, Keskin Sultan Sahin, Aktas Esin Cetin, Dogusan Zeynep, Trabzon Levent, Erdem Kuruca Dürdane Serap
Faculty of Health Sciences, Harran University, Sanliurfa, Turkey.
Department of Nanoscience and Nanoengineering, Istanbul Technical University, Istanbul, Turkey.
Biomed Microdevices. 2025 Jun 28;27(3):33. doi: 10.1007/s10544-025-00756-z.
Mononuclear cells (MNCs), a type of leukocyte, require enrichment owing to their rarity for research and clinical applications. The enrichment of MNCs is generally performed via conventional methods (e.g., density gradient centrifugation). However, these methods have downsides, such as being labor intensive, energy and time consuming, and requiring advanced equipment. Therefore, inertial microfluidics has recently drawn widespread attention as a way to overcome these limitations. This work aims to investigate MNC separation using a novel spiral inertial microfluidic system design. After MNCs were enriched by Ficoll stratification, the cells were separated according to their size and deformability properties by passing through the microfluidic system. In the final step, various cell markers were examined for characterization in these cells collected at outlets. In this paper, we determined that MNCs obtained from three different hematological products could be sorted with a recovery rate of 97.5% and a purity level of 84%, whereas red blood cells (RBCs) had a depletion ratio of 80% using Sunflower-designed microfluidic system. The loss of MNCs in this system was much lower than that in density gradient centrifugation. The separation technique studied here has several advantages, such as continuous processing, a high operation flow rate (e.g., 0.7 ml/min), simplifying the operative procedures for automation, and creating no clogging problems. Additionally, this technique can be easily integrated with downstream applications, such as direct analysis of MNCs via a flow cytometer, and can reduce the number of man-hand manipulation processes.
单核细胞（MNCs）是一种白细胞，由于其数量稀少，在研究和临床应用中需要进行富集。MNCs的富集通常通过传统方法（如密度梯度离心）来进行。然而，这些方法存在缺点，比如劳动强度大、耗费能源和时间，并且需要先进的设备。因此，惯性微流控技术最近作为一种克服这些局限性的方法受到了广泛关注。这项工作旨在研究使用一种新型螺旋惯性微流控系统设计来分离MNCs。通过Ficoll分层富集MNCs后，细胞通过微流控系统，根据其大小和可变形性进行分离。在最后一步，对在出口处收集的这些细胞中的各种细胞标志物进行检测以进行表征。在本文中，我们确定使用向日葵设计的微流控系统，从三种不同血液制品中获得的MNCs可以以97.5%的回收率和84%的纯度水平进行分选，而红细胞（RBCs）的去除率为80%。该系统中MNCs的损失远低于密度梯度离心法。这里研究的分离技术具有几个优点，例如连续处理、高操作流速（如0.7毫升/分钟）、简化自动化操作程序且不会产生堵塞问题。此外，该技术可以很容易地与下游应用集成，如通过流式细胞仪直接分析MNCs，并且可以减少人工操作过程的数量。
