SCHEDULED MAINTENANCE
Proline-conditioning and chemically-programmed ice nucleation protects spheroids during cryopreservation†
Qiongyu
Guo 
*c
and
Matthew I.
Gibson
*ab
*c
and
Matthew I.
Gibson
*ab
Abstract
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.
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