Abstract
The mass cell culture is an inevitable process for tissue engineering and regenerative medicine, because the therapies are requited a lot of cells. However, the process has not been optimized for maintaining cell activity. Especially, cell detachment using trypsin causes a damage of cells, because surface proteins of cells are degraded. Hence, a novel process to replace trypsinization has been required. In this research, the detachment method—combination of DLC-treated dish and acoustic pressure exposure is developed. The DLC-treated dish has 50 nm DLC layer and the intensity ratio of ID/IG is 0.70. In order to generate acoustic pressure, a piezoelectric ceramic plate is glued on a glass plate to configure an ultrasonic transducer. The glass plate and a polycarbonate wall compose a chamber in which a DLC-treated dish is placed via glycerol. Glycerol transmits acoustic pressure to cells that adhere on the DLC-treated dish. In the results of cell culture experiment, the number of detached cells by using proposed method is equivalent to that by using conventional method (trypsinization). Additionally, initial adhesion of cells detached by the proposed method is improved compared with the cells detached by the conventional method. Moreover, cells detached by the proposed method had surface proteins richly, while cells detached by the conventional method decreased them. The proposed method provides a cell detachment method to keep cell viability so that it may be used in future clinical applications instead of trypsinization.
