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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8594829/ https://www.ncbi.nlm.nih.gov/pubmed/34784361 http://dx.doi.org/10.1371/journal.pone.0259571 |
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author | Kilbride, Peter Meneghel, Julie Fonseca, Fernanda Morris, John |
author_facet | Kilbride, Peter Meneghel, Julie Fonseca, Fernanda Morris, John |
author_sort | Kilbride, Peter |
collection | PubMed |
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 | Online Article Text |
id | pubmed-8594829 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-85948292021-11-17 The transfer temperature from slow cooling to cryogenic storage is critical for optimal recovery of cryopreserved mammalian cells Kilbride, Peter Meneghel, Julie Fonseca, Fernanda Morris, John PLoS One Research Article 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. Public Library of Science 2021-11-16 /pmc/articles/PMC8594829/ /pubmed/34784361 http://dx.doi.org/10.1371/journal.pone.0259571 Text en © 2021 Kilbride et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Kilbride, Peter Meneghel, Julie Fonseca, Fernanda Morris, John The transfer temperature from slow cooling to cryogenic storage is critical for optimal recovery of cryopreserved mammalian cells |
title | 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_short | 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 |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8594829/ https://www.ncbi.nlm.nih.gov/pubmed/34784361 http://dx.doi.org/10.1371/journal.pone.0259571 |
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