AbstractMicrofluidic cell separation systems have emerged as attractive alternatives to traditional techniques in recent years. These systems offer the advantages of being able to handle small sample volumes and at the same time achieve highly selective separation. Conventional separation techniques, including both fluorescence-activated cell sorting (FACS) and magnetic-activated cell sorting (MACS), typically require a pre-processing incubation step to attach ligated tags (such as fluorescent dyes or magnetic beads) to cell surfaces prior to separation. These techniques are also constrained by infrastructure and high cost. Microfluidic devices with surface-immobilized adhesion molecules eliminate the need for pre-processing incubation and are a low cost alternative.We describe the selective adhesion of smooth muscle cells and endothelial cells in microfluidic devices coated with adhesion peptides. The device geometry is such that the shear stress varies linearly as a function of flow channel length, allowing simultaneous evaluation of the effects of surface chemistry and fluid shear on cell adhesion. The adhesion peptides, val-ala-pro-gly (VAPG) and arg-glu-asp-val (REDV), are known to bind selectively to smooth muscle cells and endothelial cells, respectively. These peptides were tethered to the device surface using silane chemistry and NHS-ester coupling. Cell adhesion was examined in a shear stress range of 1.3-4.0 dyn/cm2. Under these conditions, endothelial cells show significantly higher adhesion to REDV-coated devices compared to smooth muscle cells and fibroblasts. Correspondingly, smooth muscle cell adhesion in VAPG-coated devices is much greater than that of endothelial cells and fibroblasts. This selective binding behavior is also observed when mixed suspensions of the three cell types are flowed into both types of peptide-coated microfluidic devices. These results suggest that microfluidic devices coated with REDV and VAPG can be used as effective separation tools in various applications, such as tissue engineering. Specific examples of applications in cardiac and skin tissue engineering will be discussed.
Microfluidic cell sorting: a review of the advances in the separation of cells from debulking to rare cell isolation