A highly active mineral-based ice nucleating agent supports in situ cell cryopreservation in a high throughput format

Cryopreservation of biological matter in microlitre scale volumes of liquid would be useful for a range of applications. At present, it is challenging because small volumes of water tend to supercool, and deep supercooling is known to lead to poor post-thaw cell viability. Here, we show that a mineral ice nucleator can almost eliminate supercooling in 100 mu l liquid volumes during cryopreservation. This strategy of eliminating supercooling greatly enhances cell viability relative to cryopreservation protocols with uncontrolled ice nucleation. Using infrared thermography, we demonstrate a direct relationship between the extent of supercooling and post-thaw cell viability. Using a mineral nucleator delivery system, we open the door to the routine cryopreservation of mammalian cells in multiwell plates for applications such as high throughput toxicology testing of pharmaceutical products and regenerative medicine.

Automated summary

By using a mineral ice nucleator, the issue of supercooling in microlitre liquid volumes during cryopreservation can be eliminated, leading to improved cell viability. This research opens up new possibilities for large-scale cryopreservation of mammalian cells, which can be applied in fields such as high throughput toxicology testing and regenerative medicine.