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Updated: Sep 17, 2025
Microfluidic Buffer Exchange for Interference-free Micro/Nanoparticle Cell Engineering
Published on: July 10, 2016
Separation of mononuclear cells from progenitor products by a novel inertial microfluidic method
Nilgün Okşak1, Sultan Sahin Keskin2,3, Esin Cetin Aktas4
1Faculty of Health Sciences, Harran University, Sanliurfa, Turkey. nilgunoksak@harran.edu.tr.
Abstract:
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.
Insights
This study introduces a novel spiral microfluidic system for efficient mononuclear cell (MNC) enrichment. The system achieves high purity and recovery rates, offering a faster, automated alternative to traditional methods.
Area of Science:
- Biomedical Engineering
- Cellular Biology
- Microfluidics
Background:
- Mononuclear cells (MNCs) are crucial for research and clinical applications but are rare and require enrichment.
- Conventional MNC enrichment methods like density gradient centrifugation are labor-intensive, time-consuming, and require specialized equipment.
- Inertial microfluidics presents a promising alternative to overcome the limitations of traditional cell separation techniques.
Purpose of the Study:
- To investigate the separation of mononuclear cells (MNCs) using a novel spiral inertial microfluidic system.
- To evaluate the efficiency of this microfluidic system in enriching MNCs from hematological products.
- To compare the performance of the microfluidic system with conventional density gradient centrifugation.
Main Methods:
- Enrichment of MNCs was performed using Ficoll stratification.
- Cells were subsequently separated based on size and deformability properties within a novel spiral inertial microfluidic system.
- Characterization of collected cells at different outlets was conducted using various cell markers and flow cytometry.
Main Results:
- The Sunflower-designed microfluidic system achieved an MNC recovery rate of 97.5% and a purity level of 84%.
- Red blood cell (RBC) depletion reached 80%, significantly reducing contamination.
- The microfluidic system demonstrated lower MNC loss compared to density gradient centrifugation.
Conclusions:
- The novel spiral inertial microfluidic system offers an efficient, high-throughput method for MNC separation.
- This technique provides advantages such as continuous processing, automation compatibility, and reduced manual manipulation.
- The system's ability to integrate with downstream applications like flow cytometry enhances its clinical and research utility.
