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Abstract Chinese hamster ovary (CHO) cell is the most widely used mammalian cell line for commercial production of therapeutic protein. Any presence of non-viable cells in culture medium may adversely affect subsequent functionality of these proteins. Therefore, separation of non-viable cells from suspending medium is critical in biopharmaceutical and biomedical sectors. One such method termed Deterministic Lateral Displacement has already shown promising capabilities in separating cells based on the cell size difference by taking advantage of the predictable flow laminae. However, in cases where size overlaps between viable and non-viable cells are present, separation based solely on size suffers and high-resolution separation techniques are required. Dielectrophoresis, one of the most widely used nonlinear electro-kinetic mechanism, has the potential to manipulate the same size cells depending on the dielectric properties of individual cells. In this work, we demonstrated that a DLD device can be combined with a frequency-based AC electric field to perform high resolution continuous separation of non-viable CHO cells from the viable or productive cells. The behavior of the coupled DLD-DEP device is further investigated by employing numerical simulation to check the effect of geometrical parameters of the DLD arrays, velocities of the flow field and required applied voltages. A moderate row shift fraction with velocity 700μm/s provided a good separation behavior without any trapping. The cell viability was also ensured by maintaining proper electric field which otherwise may cause cell loss due to ion leakage. Our developed numerical model and presented results lay the groundwork for design and fabrication of high resolution DLD-DEP microchips for enhanced separation of viable and nonviable cells.
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Development of 2D Microfluidics Surface with Low-Frequency Electric Fields for Cell Separation Applications
Cell separation techniques are widely used in many biomedical and clinical applications for the development of screening, diagnosis and therapeutic tests. Current 3D microfluidics-based cell separation methods have limited applications in part due to low throughput and technical complexity. To address these critical needs, we have developed a 2D microfluidics surface which is the miniaturized version of a 3D microfluids cell separation device. Using low-frequency electric fields (1–10 Vpp and 1 kHz–20 MHz), we have first studied dielectrophoresis, AC electro-osmosis and capillary flow within a sessile drop, and finally utilized the results to develop the 2D cell separation surface. Our study has demonstrated that frequency-dependent dielectrophoretic force and AC electro-osmotic flow can be integrated to minimize the capillary flow and subsequently produce clusters of target cells within the 2D microfluidics surface. To demonstrate the concept, we have isolated the blood cells from a red blood cell-lysed blood sample. Cell isolation results show that significant improvement in throughout up to about 120-fold over 3D microfluidics devices. Additionally, due to the technical simplicity, this device offers great potential for use in a wide range of biomedical and clinical applications.
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- PAR ID:
- 10661126
- Publisher / Repository:
- MDPI
- Date Published:
- Journal Name:
- Sensors
- Volume:
- 25
- Issue:
- 18
- ISSN:
- 1424-8220
- Page Range / eLocation ID:
- 5816
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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- Free Publicly Accessible Full Text
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Accepted Manuscript
- Journal Article:
- https://doi.org/10.3390/s25185816
