Liquidus tracking: controlled rate vitrification for the cryopreservation of larger volumes and tissues.

BACKGROUND

Vitrification of cells or tissue at controlled cooling rates suitable for larger volumes, and with reduced cryoprotectant toxicity.

OBJECTIVE

To set out the current understanding of the LiquidusTracking (LT) vitrification technique, and to discuss the challenges and benefits of translating the method into laboratory protocols more generally applicable to meet requirements of large volume and 3-D cryo-banking in the era of regenerative medicine.

METHODS

By adding small amounts of cryoprotectants at each step and subsequently cooling the sample just above its freezing point before further increasing CPA concentration, cryoprotectant toxicity is minimized.

RESULT

CPA toxicity can be reduced by lowering the temperature. Different manual approaches to LT were evaluated and further improved.

CONCLUSIONS

Manual liquidus tracking is complicated and exhibits potential high variability. Nevertheless, this approach offers the possibility of testing several conditions simultaneously and could be used to pre-test conditions prior to automatic LT development.