Development of Micro Perfusion Cell Culture Device to Create In Vivo-Like Environments for Long-Period and Real-Time Monitoring of Cells Activities

Although enormous efforts have been consumed to improve in vitro cell-based assays for life science, environmental assessment, medical applications, etc., conventional methods still have many problems, such as large dead volumes, difficulty to maintain constant concentration of nutrition and to remove accumulating waste products in static dish-based culture, etc. In order to solve these problems, it would be a breakthrough to realize miniaturized perfusion cell culture system, which continuously supply nutrition and remove metabolic waste products during cell culture. Such a system would be possible to culture cells for a long period and could monitor chemical response at real-time. This scheme of culture system could innovatively be advanced by coupling the recent microfabrication technology. Here, we have developed and experimentally demonstrated a micro cell culture device embedded with a stirrer-type micropump for on-chip perfusion of culture medium and optical fiber connectors for on-chip and real-time fluorescent detection. The cell culture chamber was separated into two compartments by a polyester-made semi-permeable membrane, which can realize polarized cell culture condition and detect a fluorescent signal movement in-between the two-compartments. As an evaluation of the device, Caco-2 cells were successfully cultured over a period up to 26 days and investigated the activity of P-glycoprotein using a fluorescent dye of Rhodamine-123 as a substrate of P-glycoprotein