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

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 cryode...

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Autores principales:	Peter Kilbride, Julie Meneghel, Fernanda Fonseca, John Morris
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/86a1790bbc2f4b638fb92ebfc6ad0c23
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spelling	oai:doaj.org-article:86a1790bbc2f4b638fb92ebfc6ad0c232021-11-25T06:13:56ZThe transfer temperature from slow cooling to cryogenic storage is critical for optimal recovery of cryopreserved mammalian cells1932-6203https://doaj.org/article/86a1790bbc2f4b638fb92ebfc6ad0c232021-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8594829/?tool=EBIhttps://doaj.org/toc/1932-6203Cryopreservation 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.Peter KilbrideJulie MeneghelFernanda FonsecaJohn MorrisPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q 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
description	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.
format	article
author	Peter Kilbride Julie Meneghel Fernanda Fonseca John Morris
author_facet	Peter Kilbride Julie Meneghel Fernanda Fonseca John Morris
author_sort	Peter Kilbride
title	The transfer temperature from slow cooling to cryogenic storage is critical for optimal recovery of cryopreserved mammalian cells
title_short	The transfer temperature from slow cooling to cryogenic storage is critical for optimal recovery of cryopreserved mammalian cells
title_full	The transfer temperature from slow cooling to cryogenic storage is critical for optimal recovery of cryopreserved mammalian cells
title_fullStr	The transfer temperature from slow cooling to cryogenic storage is critical for optimal recovery of cryopreserved mammalian cells
title_full_unstemmed	The transfer temperature from slow cooling to cryogenic storage is critical for optimal recovery of cryopreserved mammalian cells
title_sort	transfer temperature from slow cooling to cryogenic storage is critical for optimal recovery of cryopreserved mammalian cells
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/86a1790bbc2f4b638fb92ebfc6ad0c23
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The transfer temperature from slow cooling to cryogenic storage is critical for optimal recovery of cryopreserved mammalian cells

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