| 研究実績の概要 |
To realize hepatocytes-based assays exhibiting many biological functions, we investigated the feasibilities of microwell device and microfluidic system for better metabolism and future bile recovery. In the case of the microwell device, we made a honeycomb network of pyramidal microwells to create small aggregates. In addition, we optimized the microenvironment by the overlay of Matrigel or a liver sinusoidal endothelial cell line. Although we successfully formed stably-attached aggregates, we were unfortunately not able to control precisely the formation of bile pocket in the center of the microwell. We therefore decided to change from microwell to "virtual microwell", namely proteins patterning on PDMS surface. It is on-going at this moment. As an another approach, we changed the design of previous our devices (Nakao et al., Biomicrofluidics, 5, 22212, 2011) to increase the diffusion between the culture medium and cells. To be able to monitor several devices at the time, we made a setup of 8 parallel devices to decrease the flow rate. Cell inoculation to the culture area was successfully checked in real-time. We tried different design and conditions and finally were able to fill the culture area and to get two lines of fixed hepatocytes. However, we have not yet succeeded completely when we use freshly-isolated cells at this moment. The experiments are also on-going now.
|
