Cell Motion in a Two-Stream Microfluidic Channel

Microfluidic channels have been proposed as a method for removal of cryoprotective agents from cell suspensions [Fleming, Longmire, and Hubel, J. Biomech. Eng. 129, 703 (2007)]. The device tested consists of a rectangular cross section channel of 500 $μm$ depth, 25 mm width, and 160 mm length, through which a cell suspension and wash stream flow in parallel. Cryoprotective agents diffuse from the cell stream to the wash stream and the wash stream is discarded. The washed cell stream is then ready for use. This device must be capable of removing 95% of the dimethyl sulfoxide (DMSO) from the cell stream with minimal cell losses. Our previous studies have demonstrated our ability to remove DMSO [Mata, Longmire, McKenna, Glass, and Hubel, Microfluid. Nanofluid. 5, 529 (2008)]. The next phase of the investigation involves characterizing the influence of flow conditions on cell motion through the device. To that end, Jurkat cells (lymphoblasts) in a 10% DMSO solution were flowed through the microfluidic channel in parallel with a wash stream composed of phosphate buffered saline solution (PBS). Average cell stream velocities were varied from 0.94 to 8.5 mm/s (Re 1.7 to 6.0). Cell viability at the outlet was high, indicating that cells are not damaged during their passage through the device. Gravitational settling caused an accumulation of cells near the bottom of the channel, where flow velocities are low. Cell settling leads results in an initial transient period for cell motion through the device. For the initial portion of cells flowing through the device, cells tend to accumulate in the device until a critical device population time is reached. Cell recovery (number of cells out of the device divided by the number of cells input to the device) is high (90–100%) after the device has been fully populated. For a single stage device with average cell stream velocities of $⩾6$ mm/s, cell recovery was 90–100%. As more stages are added to the device, the population time for the device increases. Gravitational settling of cells also leads to a time-varying cell concentration from the input syringe to the inlet of the channel, as well as cell losses due to cells remaining in the horizontally-oriented syringe. Reorienting the syringes to a vertical position eliminates these losses. Cell motion within the channel can be modulated by the flow conditions used. For sufficiently high Reynolds numbers, the Segre-Silberberg effect [Segre and Silberberg, J. Fluid Mech. 14, 115 (1962)] can be used to move cells from the low velocity region of the cell stream to a higher velocity region thereby reducing the transient portion of processing the cells and improving overall recovery of cells through the device.