Abstract
T cell migration plays an important role in inflammation and wound healing. They circulate in the blood and migrate into different tissues to execute their tasks. In this study, we investigated the effects of substrate stiffness, cell activation, diabetes mellitus (DM), and hydrostatic pressure on T cell migration. We hypothesized that stiffer substrates and application of hydrostatic pressure increase the speed of T cell migration, and that defect in T cell activation and DM condition decrease the migration speed. Mouse QL9 T cells were used and live cell images were taken to analyze the migration speed. Polyacrylamide (PA) gels of stiffness varying from 1 to 34 kPa and with graded stiffness were used to study the effects of substrate stiffness on T cell migration. Poly(dimethylsiloxane) (PDMS)-based cell chips were fabricated to apply a 10 cmH2O hydrostatic pressure to the cultured cells. The DM condition was simulated using STZ-treated mice. The T cell activation was modulated via application of interleukin-2 (IL-2) and defect of cell activation was simulated by knocking out CD28, an important cell-membrane receptor for T cell activation. Since application of the hydrostatic pressure may affect the traffic of water through cell membrane via aquaporin1 (AQP1), which has been shown to play an important role in cell migration, we also examined whether application of hydrostatic pressure facilitated the expression of AQP1. Our results indicate that the T cell migration was not sensitive to the variation of substrate stiffness. Application of a 10 cmH2O hydrostatic pressure may enhance the migration speed, but the difference was not significant when compared with cells without pressure exposure. Application of the 10 cmH2O enhanced AQP1 expression, but the relationships among AQP1 expression, migration speed, and hydrostatic pressure require further studies to clarify. DM condition and knockout of CD28 appeared to decrease the migration speed, but the difference was either not significant when compared with control.
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