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Subzero Nonfreezing Hypothermia with Insect Antifreeze Protein Dramatically Improves Survival Rate of Mammalian Cells

Cells for therapeutic use are often preserved at +4 °C, and the storage period is generally limited to 2–3 days. Here, we report that the survival rate (%) of mammalian cells is improved to 10–20 days when they are preserved with a subzero supercooled solution containing the antifreeze protein (AFP)...

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Autores principales: Yamauchi, Akari, Miura, Ai, Kondo, Hidemasa, Arai, Tatsuya, Sasaki, Yuji C., Tsuda, Sakae
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8657916/
https://www.ncbi.nlm.nih.gov/pubmed/34884483
http://dx.doi.org/10.3390/ijms222312680
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author Yamauchi, Akari
Miura, Ai
Kondo, Hidemasa
Arai, Tatsuya
Sasaki, Yuji C.
Tsuda, Sakae
author_facet Yamauchi, Akari
Miura, Ai
Kondo, Hidemasa
Arai, Tatsuya
Sasaki, Yuji C.
Tsuda, Sakae
author_sort Yamauchi, Akari
collection PubMed
description Cells for therapeutic use are often preserved at +4 °C, and the storage period is generally limited to 2–3 days. Here, we report that the survival rate (%) of mammalian cells is improved to 10–20 days when they are preserved with a subzero supercooled solution containing the antifreeze protein (AFP), for which an ability to stabilize both supercooled water and cell membrane integrity has been postulated. We chose adherent rat insulinoma (RIN-5F) cells as the preservation target, which were immersed into −5 °C-, −2 °C-, or +4 °C-chilled “unfrozen” solution of Euro-Collins or University of Washington (UW) containing the AFP sample obtained from insect or fish. Our results show that the survival rate of the cells preserved with the solution containing insect AFP was always higher than that of the fish AFP solution. A combination of the −5 °C-supercooling and insect AFP gave the best preservation result, namely, UW solution containing insect AFP kept 53% of the cells alive, even after 20 days of preservation at −5 °C. The insect AFP locates highly organized ice-like waters on its molecular surface. Such waters may bind to semiclathrate waters constructing both embryonic ice crystals and a membrane–water interface in the supercooled solution, thereby protecting the cells from damage due to chilling.
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spelling pubmed-86579162021-12-10 Subzero Nonfreezing Hypothermia with Insect Antifreeze Protein Dramatically Improves Survival Rate of Mammalian Cells Yamauchi, Akari Miura, Ai Kondo, Hidemasa Arai, Tatsuya Sasaki, Yuji C. Tsuda, Sakae Int J Mol Sci Article Cells for therapeutic use are often preserved at +4 °C, and the storage period is generally limited to 2–3 days. Here, we report that the survival rate (%) of mammalian cells is improved to 10–20 days when they are preserved with a subzero supercooled solution containing the antifreeze protein (AFP), for which an ability to stabilize both supercooled water and cell membrane integrity has been postulated. We chose adherent rat insulinoma (RIN-5F) cells as the preservation target, which were immersed into −5 °C-, −2 °C-, or +4 °C-chilled “unfrozen” solution of Euro-Collins or University of Washington (UW) containing the AFP sample obtained from insect or fish. Our results show that the survival rate of the cells preserved with the solution containing insect AFP was always higher than that of the fish AFP solution. A combination of the −5 °C-supercooling and insect AFP gave the best preservation result, namely, UW solution containing insect AFP kept 53% of the cells alive, even after 20 days of preservation at −5 °C. The insect AFP locates highly organized ice-like waters on its molecular surface. Such waters may bind to semiclathrate waters constructing both embryonic ice crystals and a membrane–water interface in the supercooled solution, thereby protecting the cells from damage due to chilling. MDPI 2021-11-24 /pmc/articles/PMC8657916/ /pubmed/34884483 http://dx.doi.org/10.3390/ijms222312680 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Yamauchi, Akari
Miura, Ai
Kondo, Hidemasa
Arai, Tatsuya
Sasaki, Yuji C.
Tsuda, Sakae
Subzero Nonfreezing Hypothermia with Insect Antifreeze Protein Dramatically Improves Survival Rate of Mammalian Cells
title Subzero Nonfreezing Hypothermia with Insect Antifreeze Protein Dramatically Improves Survival Rate of Mammalian Cells
title_full Subzero Nonfreezing Hypothermia with Insect Antifreeze Protein Dramatically Improves Survival Rate of Mammalian Cells
title_fullStr Subzero Nonfreezing Hypothermia with Insect Antifreeze Protein Dramatically Improves Survival Rate of Mammalian Cells
title_full_unstemmed Subzero Nonfreezing Hypothermia with Insect Antifreeze Protein Dramatically Improves Survival Rate of Mammalian Cells
title_short Subzero Nonfreezing Hypothermia with Insect Antifreeze Protein Dramatically Improves Survival Rate of Mammalian Cells
title_sort subzero nonfreezing hypothermia with insect antifreeze protein dramatically improves survival rate of mammalian cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8657916/
https://www.ncbi.nlm.nih.gov/pubmed/34884483
http://dx.doi.org/10.3390/ijms222312680
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