マイクロ流体プローブ集積型デバイスを用いた局所刺激に対する細胞応答評価

抄録

In this study, we have developed a novel microfluidic device to control a local chemical stimulation area in cell culture spaces toward cell-based assays. The device consists of a cell culture channel and a microfluidic probe (MFP) channel which has a pair of the injection and aspiration channels installed on the ceiling of the cell culture chamber. A local stimulation area within the cell chamber can be controlled by the flow ratio of injection/aspiration channels. Also, it is possible to culture cells under perfusion by aspirating the culture medium from the reservoir using a syringe pump. To achieve precise fluid handling, the device was installed into the jig as an interface to syringe pumps. The device has been evaluated by fluidic simulation using Finite Element Method (FEM), a flow control experiment and chemical stimulation experiment on cells. The result showed that control of chemical stimulation from several cells to cell population scale in the chamber can be possible by regulating the flow ratio. The local chemical stimulation area enables grasped by fluidic simulation because the result of the experiment and simulation are approximate in case of flow control experiment and cell staining experiment. In addition, Ca²⁺ propagation in response to chemical factors was observed using the device. We conclude that the device can be powerful tool for biological applications such as the differentiation control of pluripotent stem cells and other cells signaling studies.