Freeze-drying of mammalian cells using trehalose: preservation of DNA integrity

Abstract The aim of this study was to investigate preservation of biomolecular structures, particularly DNA, in freeze-dried fibroblasts, after loading with trehalose via freezing-induced uptake. Cells were freeze-dried with trehalose alone or in a mixture of albumin and trehalose. Albumin was added...

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Autores principales:	Miao Zhang, Harriëtte Oldenhof, Bulat Sydykov, Judith Bigalk, Harald Sieme, Willem F. Wolkers
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6818fc6151594e72a6ebc0ad9c7eefd8
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spelling	oai:doaj.org-article:6818fc6151594e72a6ebc0ad9c7eefd82021-12-02T16:07:56ZFreeze-drying of mammalian cells using trehalose: preservation of DNA integrity10.1038/s41598-017-06542-z2045-2322https://doaj.org/article/6818fc6151594e72a6ebc0ad9c7eefd82017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-06542-zhttps://doaj.org/toc/2045-2322Abstract The aim of this study was to investigate preservation of biomolecular structures, particularly DNA, in freeze-dried fibroblasts, after loading with trehalose via freezing-induced uptake. Cells were freeze-dried with trehalose alone or in a mixture of albumin and trehalose. Albumin was added to increase the glass transition temperature and storage stability. No viable cells were recovered after freeze-drying and rehydration. FTIR studies showed that membrane phase behavior of freeze-dried cells resembles that of fresh cells. However, one day after rehydration membrane phase separation was observed, irrespective of the presence or absence of trehalose during freeze-drying. Freeze-drying did not affect the overall protein secondary structure. Analysis of DNA damage via single cell gel electrophoresis (‘comet assay’) showed that DNA damage progressively increased with storage duration and temperature. DNA damage was prevented during storage at 4 °C. It is shown that trehalose reduces DNA damage during storage, whereas addition of albumin did not seem to have an additional protective effect on storage stability (i.e. DNA integrity) despite the fact that albumin increased the glass transition temperature. Taken together, DNA in freeze-dried somatic cells can be preserved using trehalose as protectant and storage at or below 4 °C.Miao ZhangHarriëtte OldenhofBulat SydykovJudith BigalkHarald SiemeWillem F. WolkersNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-10 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Miao Zhang Harriëtte Oldenhof Bulat Sydykov Judith Bigalk Harald Sieme Willem F. Wolkers Freeze-drying of mammalian cells using trehalose: preservation of DNA integrity
description	Abstract The aim of this study was to investigate preservation of biomolecular structures, particularly DNA, in freeze-dried fibroblasts, after loading with trehalose via freezing-induced uptake. Cells were freeze-dried with trehalose alone or in a mixture of albumin and trehalose. Albumin was added to increase the glass transition temperature and storage stability. No viable cells were recovered after freeze-drying and rehydration. FTIR studies showed that membrane phase behavior of freeze-dried cells resembles that of fresh cells. However, one day after rehydration membrane phase separation was observed, irrespective of the presence or absence of trehalose during freeze-drying. Freeze-drying did not affect the overall protein secondary structure. Analysis of DNA damage via single cell gel electrophoresis (‘comet assay’) showed that DNA damage progressively increased with storage duration and temperature. DNA damage was prevented during storage at 4 °C. It is shown that trehalose reduces DNA damage during storage, whereas addition of albumin did not seem to have an additional protective effect on storage stability (i.e. DNA integrity) despite the fact that albumin increased the glass transition temperature. Taken together, DNA in freeze-dried somatic cells can be preserved using trehalose as protectant and storage at or below 4 °C.
format	article
author	Miao Zhang Harriëtte Oldenhof Bulat Sydykov Judith Bigalk Harald Sieme Willem F. Wolkers
author_facet	Miao Zhang Harriëtte Oldenhof Bulat Sydykov Judith Bigalk Harald Sieme Willem F. Wolkers
author_sort	Miao Zhang
title	Freeze-drying of mammalian cells using trehalose: preservation of DNA integrity
title_short	Freeze-drying of mammalian cells using trehalose: preservation of DNA integrity
title_full	Freeze-drying of mammalian cells using trehalose: preservation of DNA integrity
title_fullStr	Freeze-drying of mammalian cells using trehalose: preservation of DNA integrity
title_full_unstemmed	Freeze-drying of mammalian cells using trehalose: preservation of DNA integrity
title_sort	freeze-drying of mammalian cells using trehalose: preservation of dna integrity
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/6818fc6151594e72a6ebc0ad9c7eefd8
work_keys_str_mv	AT miaozhang freezedryingofmammaliancellsusingtrehalosepreservationofdnaintegrity AT harrietteoldenhof freezedryingofmammaliancellsusingtrehalosepreservationofdnaintegrity AT bulatsydykov freezedryingofmammaliancellsusingtrehalosepreservationofdnaintegrity AT judithbigalk freezedryingofmammaliancellsusingtrehalosepreservationofdnaintegrity AT haraldsieme freezedryingofmammaliancellsusingtrehalosepreservationofdnaintegrity AT willemfwolkers freezedryingofmammaliancellsusingtrehalosepreservationofdnaintegrity
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Freeze-drying of mammalian cells using trehalose: preservation of DNA integrity

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