2021 Volume 67 Issue 1 Pages 31-40
Supercool preservation method for biological tissues has attracted attention, because the supercooled state can reduce physical damage of living cells and tissues. There are many reports that living tissues could be preserved in a solution containing cryoprotectant at a supercooled state. However, the solution with cryoprotectant has risks of cytotoxicity for living cells. Therefore, the supercool preservation method using a solution with lower cytotoxicity is required. In this study, we focused on the supercool preservation using a cell culture medium without cryoprotectant for living cell and tissue preservation. For a fundamental study, the living cells suspended in the culture medium without cryoprotectant were subjected to the supercooling condition to elucidate the effect of supercooling preservation on cell viability and proliferation. Specifically, a custom-made supercooling device to preserve cell-suspended solution was developed. Human neonatal fibroblasts were suspended in culture medium and stored inside the device at -4°C and -8°C of supercooling state for 24 hours. As a result, there was no significant difference in the survival rate after supercool preservation due to the difference in the supercooling temperature. On the other hand, both cell adhesion and cell proliferation rate were higher in a supercooled state at -4°C than stored at -8°C.
