Gene expression analysis of human induced pluripotent stem cells cryopreserved by vitrification using StemCell Keep

In recent years, regenerative medicine research using human somatic and induced pluripotent stem cells has advanced considerably, promoting clinical applications. However, it is essential that these cells are cryopreserved safely and effectively. Most cryopreservation solution agents contain dimethy...

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Autores principales:	Akemi Ota, Suong-Hyu Hyon, Shoichiro Sumi, Kazuaki Matsumura
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Human induced pluripotent stem cell Cryopreservation Vitrification DNA microarray Gene enrichment analysis Biology (General) QH301-705.5 Biochemistry QD415-436
Acceso en línea:	https://doaj.org/article/53fd777e0a5645b8993d660fc84465cd
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spelling	oai:doaj.org-article:53fd777e0a5645b8993d660fc84465cd2021-11-18T04:50:31ZGene expression analysis of human induced pluripotent stem cells cryopreserved by vitrification using StemCell Keep2405-580810.1016/j.bbrep.2021.101172https://doaj.org/article/53fd777e0a5645b8993d660fc84465cd2021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2405580821002661https://doaj.org/toc/2405-5808In recent years, regenerative medicine research using human somatic and induced pluripotent stem cells has advanced considerably, promoting clinical applications. However, it is essential that these cells are cryopreserved safely and effectively. Most cryopreservation solution agents contain dimethyl sulfoxide (DMSO), which exhibits strong toxicity and can potentially promote cell differentiation. Hence, it is important to explore substitutes for DMSO in cryoprotectant solutions. One such alternative is StemCell Keep (SCK), a DMSO-free solution that has been reported to effectively cryopreserve human induced pluripotent stem cells (hiPS cells). To clarify the effect of cryopreservation agents on cells, DNA microarray analysis is useful, as it can identify a large number of gene expression differences in cryopreserved cells, as well as functional increases in gene groups. In this study, we performed gene expression analysis of SCK-cryopreserved hiPS cells using a DNA microarray gene chip. The hiPS cells vitrified with SCK or DMSO-based vitrification solutions were thawed and cultured on Matrigel under feeder-free conditions, and RNA was extracted for DNA microarray analysis. Genes obtained from DNA microarray data were classified by the keywords of Gene Ontology Biological Process Term, and their relationships were analyzed using DAVID or the GeneMANIA database.SCK-cryopreserved hiPS cells expressed several anti-apoptotic genes, as well as genes related to cell adhesion or proliferation at levels that were nearly equivalent to those of non-frozen hiPS cells. Gene enrichment analysis with selected genes of SCK-cryopreserved hiPS cells whose expression differences were superior to those of DAP-cryopreserved showed strong interactions of negative regulation of apoptotic process, cell adhesion and positive regulation of cell proliferation in DAVID analysis. We demonstrated that SCK successfully maintained the key functions of hiPS cells, including anti-apoptosis, cell adhesion, and cell proliferation, during cryopreservation.Akemi OtaSuong-Hyu HyonShoichiro SumiKazuaki MatsumuraElsevierarticleHuman induced pluripotent stem cellCryopreservationVitrificationDNA microarrayGene enrichment analysisBiology (General)QH301-705.5BiochemistryQD415-436ENBiochemistry and Biophysics Reports, Vol 28, Iss , Pp 101172- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Human induced pluripotent stem cell Cryopreservation Vitrification DNA microarray Gene enrichment analysis Biology (General) QH301-705.5 Biochemistry QD415-436
spellingShingle	Human induced pluripotent stem cell Cryopreservation Vitrification DNA microarray Gene enrichment analysis Biology (General) QH301-705.5 Biochemistry QD415-436 Akemi Ota Suong-Hyu Hyon Shoichiro Sumi Kazuaki Matsumura Gene expression analysis of human induced pluripotent stem cells cryopreserved by vitrification using StemCell Keep
description	In recent years, regenerative medicine research using human somatic and induced pluripotent stem cells has advanced considerably, promoting clinical applications. However, it is essential that these cells are cryopreserved safely and effectively. Most cryopreservation solution agents contain dimethyl sulfoxide (DMSO), which exhibits strong toxicity and can potentially promote cell differentiation. Hence, it is important to explore substitutes for DMSO in cryoprotectant solutions. One such alternative is StemCell Keep (SCK), a DMSO-free solution that has been reported to effectively cryopreserve human induced pluripotent stem cells (hiPS cells). To clarify the effect of cryopreservation agents on cells, DNA microarray analysis is useful, as it can identify a large number of gene expression differences in cryopreserved cells, as well as functional increases in gene groups. In this study, we performed gene expression analysis of SCK-cryopreserved hiPS cells using a DNA microarray gene chip. The hiPS cells vitrified with SCK or DMSO-based vitrification solutions were thawed and cultured on Matrigel under feeder-free conditions, and RNA was extracted for DNA microarray analysis. Genes obtained from DNA microarray data were classified by the keywords of Gene Ontology Biological Process Term, and their relationships were analyzed using DAVID or the GeneMANIA database.SCK-cryopreserved hiPS cells expressed several anti-apoptotic genes, as well as genes related to cell adhesion or proliferation at levels that were nearly equivalent to those of non-frozen hiPS cells. Gene enrichment analysis with selected genes of SCK-cryopreserved hiPS cells whose expression differences were superior to those of DAP-cryopreserved showed strong interactions of negative regulation of apoptotic process, cell adhesion and positive regulation of cell proliferation in DAVID analysis. We demonstrated that SCK successfully maintained the key functions of hiPS cells, including anti-apoptosis, cell adhesion, and cell proliferation, during cryopreservation.
format	article
author	Akemi Ota Suong-Hyu Hyon Shoichiro Sumi Kazuaki Matsumura
author_facet	Akemi Ota Suong-Hyu Hyon Shoichiro Sumi Kazuaki Matsumura
author_sort	Akemi Ota
title	Gene expression analysis of human induced pluripotent stem cells cryopreserved by vitrification using StemCell Keep
title_short	Gene expression analysis of human induced pluripotent stem cells cryopreserved by vitrification using StemCell Keep
title_full	Gene expression analysis of human induced pluripotent stem cells cryopreserved by vitrification using StemCell Keep
title_fullStr	Gene expression analysis of human induced pluripotent stem cells cryopreserved by vitrification using StemCell Keep
title_full_unstemmed	Gene expression analysis of human induced pluripotent stem cells cryopreserved by vitrification using StemCell Keep
title_sort	gene expression analysis of human induced pluripotent stem cells cryopreserved by vitrification using stemcell keep
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/53fd777e0a5645b8993d660fc84465cd
work_keys_str_mv	AT akemiota geneexpressionanalysisofhumaninducedpluripotentstemcellscryopreservedbyvitrificationusingstemcellkeep AT suonghyuhyon geneexpressionanalysisofhumaninducedpluripotentstemcellscryopreservedbyvitrificationusingstemcellkeep AT shoichirosumi geneexpressionanalysisofhumaninducedpluripotentstemcellscryopreservedbyvitrificationusingstemcellkeep AT kazuakimatsumura geneexpressionanalysisofhumaninducedpluripotentstemcellscryopreservedbyvitrificationusingstemcellkeep
_version_	1718424972695699456

Gene expression analysis of human induced pluripotent stem cells cryopreserved by vitrification using StemCell Keep

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