pH-Dependent migratory behaviors of neutrophil-like cells in a microfluidic device with controllability of dissolved gas concentrations

Inflammatory microenvironments often become acidic (pH < 7.4) due to tissue oxygen deprivation and lactate release in glycolysis by activated immune cells. Although neutrophils are known to accumulate in such microenvironments, the effects of pH on their migration are not fully understood. Here, we first investigated the pH control around cultured cells with a microfluidic device, which was equipped with two gas channels above three parallel media channels. By supplying gas mixtures with predefined carbon dioxide (CO2) concentrations to the gas channels, the gas exchange adjusted the dissolved CO2 and affected the chemical equilibrium of sodium hydrogen carbonate in the cell culture medium. A pH gradient from 8.3 to 6.8 was generated along the media channels when gas mixtures containing 1% and 50% CO2 were supplied to the left and right gas channels, respectively. Neutrophil-like differentiated human promyelocytic leukemia cells (HL-60) were then seeded to the fibronectin-coated media channels and their migratory behaviors were quantified while varying the pH. The cell migration became more active and faster under high pH than under low pH conditions. However, no directional migration along the pH gradient was detected during the three-hour observation. Thus, the microfluidic device is useful to elucidate pH-dependent cellular dynamics.