Proline-conditioning and chemically-programmed ice nucleation protects spheroids during cryopreservation

Spheroids mimic 3-D tissue niches better than standard cell cultures. Cryopreserving spheroids, however, remains challenging as conventional cryoprotectants do not mitigate all damage mechanisms. Here chemically-programmed extracellular ice nucleation is used to prevent supercooling, alongside proline pre-conditioning, which are found to synergystically improve post-thaw recovery of spheroids. This validates the need to identify compounds and materials to address both biochemical and biophysical damage pathways beyond standard cryoprotectants.

Automated summary

Spheroids are better at mimicking 3-D tissue niches than standard cell cultures, but cryopreserving them is challenging due to the limitations of conventional cryoprotectants. This study demonstrates that chemically-programmed extracellular ice nucleation combined with proline pre-conditioning can significantly improve post-thaw recovery of spheroids. This highlights the importance of identifying compounds and materials to address both biochemical and biophysical damage pathways beyond standard cryoprotectants.