Cell culture and variations in the comet assay do not affect the genomic integrity of adipose-derived stem cells
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Acta Scientiarum Biological Sciences |
Texto Completo: | http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/60714 |
Resumo: | The application of quality control tests, such as the comet assay, are essential when adipose-derived stem cells are cultured for therapeutic purposes. However, the steps involved in the development of this assay should be investigated, in order to reduce their influence on genomic damage in cells. The study aimed to evaluate if the cell culture process causes DNA damage, and if variations in the lysis time and pH of the electrophoresis buffer interfere in the genotoxicity results. Four different comet assay protocols were evaluated, and the effects of lysis time and pH conditions of the electrophoresis buffer solution were stated as follows: 2 hours and pH 12; 24 hours and pH 12; 2 hours and pH ≥ 13 and 24 hours and pH ≥ 13. The tail moment was analyzed, and results indicated that at the time cells were detached from the flasks, there was little damage to the DNA in the adipose-derived stem cells, which was confirmed by evaluation of the expression of mRNA genes involved in damage and repair processes of genetic material. Also, the tail moment values did not show significant differences among the four evaluated protocols (p < 0.05), with no indication of damage when compared to the positive control (p < 0.05). Thus, any of the tested protocols can be applied in genotoxicity tests with adipose-derived stem cells, without causing damage to them. |
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Cell culture and variations in the comet assay do not affect the genomic integrity of adipose-derived stem cellsCell culture and variations in the comet assay do not affect the genomic integrity of adipose-derived stem cellsDNA damage; gene expression; polymerase chain reaction.DNA damage; gene expression; polymerase chain reaction.The application of quality control tests, such as the comet assay, are essential when adipose-derived stem cells are cultured for therapeutic purposes. However, the steps involved in the development of this assay should be investigated, in order to reduce their influence on genomic damage in cells. The study aimed to evaluate if the cell culture process causes DNA damage, and if variations in the lysis time and pH of the electrophoresis buffer interfere in the genotoxicity results. Four different comet assay protocols were evaluated, and the effects of lysis time and pH conditions of the electrophoresis buffer solution were stated as follows: 2 hours and pH 12; 24 hours and pH 12; 2 hours and pH ≥ 13 and 24 hours and pH ≥ 13. The tail moment was analyzed, and results indicated that at the time cells were detached from the flasks, there was little damage to the DNA in the adipose-derived stem cells, which was confirmed by evaluation of the expression of mRNA genes involved in damage and repair processes of genetic material. Also, the tail moment values did not show significant differences among the four evaluated protocols (p < 0.05), with no indication of damage when compared to the positive control (p < 0.05). Thus, any of the tested protocols can be applied in genotoxicity tests with adipose-derived stem cells, without causing damage to them.The application of quality control tests, such as the comet assay, are essential when adipose-derived stem cells are cultured for therapeutic purposes. However, the steps involved in the development of this assay should be investigated, in order to reduce their influence on genomic damage in cells. The study aimed to evaluate if the cell culture process causes DNA damage, and if variations in the lysis time and pH of the electrophoresis buffer interfere in the genotoxicity results. Four different comet assay protocols were evaluated, and the effects of lysis time and pH conditions of the electrophoresis buffer solution were stated as follows: 2 hours and pH 12; 24 hours and pH 12; 2 hours and pH ≥ 13 and 24 hours and pH ≥ 13. The tail moment was analyzed, and results indicated that at the time cells were detached from the flasks, there was little damage to the DNA in the adipose-derived stem cells, which was confirmed by evaluation of the expression of mRNA genes involved in damage and repair processes of genetic material. Also, the tail moment values did not show significant differences among the four evaluated protocols (p < 0.05), with no indication of damage when compared to the positive control (p < 0.05). Thus, any of the tested protocols can be applied in genotoxicity tests with adipose-derived stem cells, without causing damage to them.Universidade Estadual De Maringá2022-07-28info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/6071410.4025/actascibiolsci.v44i1.60714Acta Scientiarum. Biological Sciences; Vol 44 (2022): Publicação contínua; e60714Acta Scientiarum. Biological Sciences; v. 44 (2022): Publicação contínua; e607141807-863X1679-9283reponame:Acta Scientiarum Biological Sciencesinstname:Universidade Estadual de Maringá (UEM)instacron:UEMenghttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/60714/751375154605Copyright (c) 2022 Acta Scientiarum. Biological Scienceshttp://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessBernardi, LuanaSchweich-Adami, Laynna de Carvalho Oliveira, Edwin José Torres de Baranoski, Adrivânio Oliveira, Rodrigo Juliano Silva, Andreia Conceição Milan Brochado Antoniolli da 2022-08-22T16:54:15Zoai:periodicos.uem.br/ojs:article/60714Revistahttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSciPUBhttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/oai||actabiol@uem.br1807-863X1679-9283opendoar:2022-08-22T16:54:15Acta Scientiarum Biological Sciences - Universidade Estadual de Maringá (UEM)false |
dc.title.none.fl_str_mv |
Cell culture and variations in the comet assay do not affect the genomic integrity of adipose-derived stem cells Cell culture and variations in the comet assay do not affect the genomic integrity of adipose-derived stem cells |
title |
Cell culture and variations in the comet assay do not affect the genomic integrity of adipose-derived stem cells |
spellingShingle |
Cell culture and variations in the comet assay do not affect the genomic integrity of adipose-derived stem cells Bernardi, Luana DNA damage; gene expression; polymerase chain reaction. DNA damage; gene expression; polymerase chain reaction. |
title_short |
Cell culture and variations in the comet assay do not affect the genomic integrity of adipose-derived stem cells |
title_full |
Cell culture and variations in the comet assay do not affect the genomic integrity of adipose-derived stem cells |
title_fullStr |
Cell culture and variations in the comet assay do not affect the genomic integrity of adipose-derived stem cells |
title_full_unstemmed |
Cell culture and variations in the comet assay do not affect the genomic integrity of adipose-derived stem cells |
title_sort |
Cell culture and variations in the comet assay do not affect the genomic integrity of adipose-derived stem cells |
author |
Bernardi, Luana |
author_facet |
Bernardi, Luana Schweich-Adami, Laynna de Carvalho Oliveira, Edwin José Torres de Baranoski, Adrivânio Oliveira, Rodrigo Juliano Silva, Andreia Conceição Milan Brochado Antoniolli da |
author_role |
author |
author2 |
Schweich-Adami, Laynna de Carvalho Oliveira, Edwin José Torres de Baranoski, Adrivânio Oliveira, Rodrigo Juliano Silva, Andreia Conceição Milan Brochado Antoniolli da |
author2_role |
author author author author author |
dc.contributor.author.fl_str_mv |
Bernardi, Luana Schweich-Adami, Laynna de Carvalho Oliveira, Edwin José Torres de Baranoski, Adrivânio Oliveira, Rodrigo Juliano Silva, Andreia Conceição Milan Brochado Antoniolli da |
dc.subject.por.fl_str_mv |
DNA damage; gene expression; polymerase chain reaction. DNA damage; gene expression; polymerase chain reaction. |
topic |
DNA damage; gene expression; polymerase chain reaction. DNA damage; gene expression; polymerase chain reaction. |
description |
The application of quality control tests, such as the comet assay, are essential when adipose-derived stem cells are cultured for therapeutic purposes. However, the steps involved in the development of this assay should be investigated, in order to reduce their influence on genomic damage in cells. The study aimed to evaluate if the cell culture process causes DNA damage, and if variations in the lysis time and pH of the electrophoresis buffer interfere in the genotoxicity results. Four different comet assay protocols were evaluated, and the effects of lysis time and pH conditions of the electrophoresis buffer solution were stated as follows: 2 hours and pH 12; 24 hours and pH 12; 2 hours and pH ≥ 13 and 24 hours and pH ≥ 13. The tail moment was analyzed, and results indicated that at the time cells were detached from the flasks, there was little damage to the DNA in the adipose-derived stem cells, which was confirmed by evaluation of the expression of mRNA genes involved in damage and repair processes of genetic material. Also, the tail moment values did not show significant differences among the four evaluated protocols (p < 0.05), with no indication of damage when compared to the positive control (p < 0.05). Thus, any of the tested protocols can be applied in genotoxicity tests with adipose-derived stem cells, without causing damage to them. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-07-28 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/60714 10.4025/actascibiolsci.v44i1.60714 |
url |
http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/60714 |
identifier_str_mv |
10.4025/actascibiolsci.v44i1.60714 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/60714/751375154605 |
dc.rights.driver.fl_str_mv |
Copyright (c) 2022 Acta Scientiarum. Biological Sciences http://creativecommons.org/licenses/by/4.0 info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Copyright (c) 2022 Acta Scientiarum. Biological Sciences http://creativecommons.org/licenses/by/4.0 |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Estadual De Maringá |
publisher.none.fl_str_mv |
Universidade Estadual De Maringá |
dc.source.none.fl_str_mv |
Acta Scientiarum. Biological Sciences; Vol 44 (2022): Publicação contínua; e60714 Acta Scientiarum. Biological Sciences; v. 44 (2022): Publicação contínua; e60714 1807-863X 1679-9283 reponame:Acta Scientiarum Biological Sciences instname:Universidade Estadual de Maringá (UEM) instacron:UEM |
instname_str |
Universidade Estadual de Maringá (UEM) |
instacron_str |
UEM |
institution |
UEM |
reponame_str |
Acta Scientiarum Biological Sciences |
collection |
Acta Scientiarum Biological Sciences |
repository.name.fl_str_mv |
Acta Scientiarum Biological Sciences - Universidade Estadual de Maringá (UEM) |
repository.mail.fl_str_mv |
||actabiol@uem.br |
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1799317391393423360 |