The transfer temperature from slow cooling to cryogenic storage is critical for optimal recovery of cryopreserved mammalian cells - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue PLoS ONE Année : 2021

The transfer temperature from slow cooling to cryogenic storage is critical for optimal recovery of cryopreserved mammalian cells

Peter Kilbride
  • Fonction : Auteur
  • PersonId : 1139234
Julie Meneghel
John Morris

Résumé

Cryopreservation is a key step for the effective delivery of many cell therapies and for the maintenance of biological materials for research. The preservation process must be carefully controlled to ensure maximum, post-thaw recovery using cooling rates slow enough to allow time for cells to cryodehydrate sufficiently to avoid lethal intracellular ice. This study focuses on determining the temperature necessary at the end of controlled slow cooling before transfer to cryogenic storage which ensures optimal recovery of the processed cell samples. Using nucleated, mammalian cell lines derived from liver (HepG2), ovary (CHO) and bone tissue (MG63) this study has shown that cooling must be controlled to-40˚C before transfer to long term storage to ensure optimal cell recovery. No further advantage was seen by controlling cooling to lower temperatures. These results are consistent with collected differential scanning calorimetry data, that indicated the cells underwent an intracellular, colloidal glass transition between-49 and-59˚C (Tg'i) in the presence of the cryoprotective agent dimethyl sulfoxide (DMSO). The glass forms at the point of maximum cryodehydration and no further cellular dehydration is possible. At this point the risk of lethal intracellular ice forming on transfer to ultra-low temperature storage is eliminated. In practice it may not be necessary to continue slow cooling to below this temperature as optimal recovery at-40˚C indicates that the cells have become sufficiently dehydrated to avoid further, significant damage when transferred into ultra-low temperature storage.

Mots clés

Fichier principal
Vignette du fichier
KilbrideMeneghel_2021_The transfer temperature from slow cooling to cryogenic storage is critical for optimal recovery of cryopreserved mammalian cells.pdf (1.19 Mo) Télécharger le fichier
Origine : Fichiers éditeurs autorisés sur une archive ouverte

Dates et versions

hal-03446319 , version 1 (24-11-2021)

Licence

Paternité

Identifiants

Citer

Peter Kilbride, Julie Meneghel, Fernanda Fonseca, John Morris. The transfer temperature from slow cooling to cryogenic storage is critical for optimal recovery of cryopreserved mammalian cells. PLoS ONE, 2021, 16 (11), pp.e0259571. ⟨10.1371/journal.pone.0259571⟩. ⟨hal-03446319⟩
47 Consultations
89 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More