Failure of Y-27632 to improve the culture of adult human adipose-derived stem cells

Detalhes bibliográficos
Autor(a) principal: Lamas, Nuno Jorge
Data de Publicação: 2015
Outros Autores: Serra, Sofia Cristina, Salgado, A. J., Sousa, Nuno
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/1822/34241
Resumo: Y-27632 is a well-known inhibitor of the Rho-associated coiled kinase (ROCK) and has been shown to significantly improve the culture of a variety of multipotent stem cell types. However, the effects of Y-27632 on the expansion of adult human adipose-derived stem cell (hADSC) cultures remain to be established. Here, we aimed to characterize the effects of Y-27632 on the culture of hADSCs. Adult hADSCs were isolated from subjects submitted to elective plastic surgery procedures and cultivated in vitro under optimized conditions. Our results show that the continuous supplementation of hADSC cultures with Y-27632 led to decreased numbers of cells and decreased global metabolic viability of hADSC cultures when compared with control conditions. This effect appeared to be dependent on the continuous presence of the drug and was shown to be concentration-dependent and significant for 10 muM and 20 muM of Y-27632. Moreover, the Y-27632-induced decrease in hADSC numbers was not linked to a block in global cell proliferation, as cell numbers consistently increased from the moment of plating until passaging. In addition, Y-27632 was not able to increase the number of hADSCs present in culture 24 hours after passaging. Taken together, our results suggest that, in contrast to other stem cell types, Y-27632 supplementation is not a suitable strategy to enhance hADSC culture expansion.
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spelling Failure of Y-27632 to improve the culture of adult human adipose-derived stem cellsHuman mesenchymal stem cellsHuman multipotent stromal cells (hMSCs)ROCK inhibitorMTS assayScience & TechnologyY-27632 is a well-known inhibitor of the Rho-associated coiled kinase (ROCK) and has been shown to significantly improve the culture of a variety of multipotent stem cell types. However, the effects of Y-27632 on the expansion of adult human adipose-derived stem cell (hADSC) cultures remain to be established. Here, we aimed to characterize the effects of Y-27632 on the culture of hADSCs. Adult hADSCs were isolated from subjects submitted to elective plastic surgery procedures and cultivated in vitro under optimized conditions. Our results show that the continuous supplementation of hADSC cultures with Y-27632 led to decreased numbers of cells and decreased global metabolic viability of hADSC cultures when compared with control conditions. This effect appeared to be dependent on the continuous presence of the drug and was shown to be concentration-dependent and significant for 10 muM and 20 muM of Y-27632. Moreover, the Y-27632-induced decrease in hADSC numbers was not linked to a block in global cell proliferation, as cell numbers consistently increased from the moment of plating until passaging. In addition, Y-27632 was not able to increase the number of hADSCs present in culture 24 hours after passaging. Taken together, our results suggest that, in contrast to other stem cell types, Y-27632 supplementation is not a suitable strategy to enhance hADSC culture expansion.We thank Jeffrey M Gimble (Center for Stem Cell Research and Regenerative Medicine, Tulane University and LaCell LLC, New Orleans, LA, USA) for kindly isolating, characterizing, and sharing the cellular lines of hADSCs used in the present study and for critical input on the manuscript. We also would very much like to thank Laurent Roybon (Stem Cell Laboratory for CNS Disease Modeling, Department of Experimental Medical Science, Lund University, Lund, Sweden) for the utilization of Metamorph NX software for automated cell quantification. We are grateful to Miguel Carvalho, Ana Pires, Eduardo Gomes, Fabio Teixeira and Nuno Silva for technical assistance. This work was supported by the Portuguese Foundation for Science and Technology (predoctoral fellowship to NJL [SFRH/BD/33421/2008]; FCT Investigator Program to AJS) and the Luso-American Development Foundation.Dove Press LtdUniversidade do MinhoLamas, Nuno JorgeSerra, Sofia CristinaSalgado, A. J.Sousa, Nuno2015-01-072015-01-07T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/34241eng1178-695710.2147/SCCAA.S66597http://www.dovepress.cominfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-21T12:30:13Zoai:repositorium.sdum.uminho.pt:1822/34241Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:25:20.658559Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Failure of Y-27632 to improve the culture of adult human adipose-derived stem cells
title Failure of Y-27632 to improve the culture of adult human adipose-derived stem cells
spellingShingle Failure of Y-27632 to improve the culture of adult human adipose-derived stem cells
Lamas, Nuno Jorge
Human mesenchymal stem cells
Human multipotent stromal cells (hMSCs)
ROCK inhibitor
MTS assay
Science & Technology
title_short Failure of Y-27632 to improve the culture of adult human adipose-derived stem cells
title_full Failure of Y-27632 to improve the culture of adult human adipose-derived stem cells
title_fullStr Failure of Y-27632 to improve the culture of adult human adipose-derived stem cells
title_full_unstemmed Failure of Y-27632 to improve the culture of adult human adipose-derived stem cells
title_sort Failure of Y-27632 to improve the culture of adult human adipose-derived stem cells
author Lamas, Nuno Jorge
author_facet Lamas, Nuno Jorge
Serra, Sofia Cristina
Salgado, A. J.
Sousa, Nuno
author_role author
author2 Serra, Sofia Cristina
Salgado, A. J.
Sousa, Nuno
author2_role author
author
author
dc.contributor.none.fl_str_mv Universidade do Minho
dc.contributor.author.fl_str_mv Lamas, Nuno Jorge
Serra, Sofia Cristina
Salgado, A. J.
Sousa, Nuno
dc.subject.por.fl_str_mv Human mesenchymal stem cells
Human multipotent stromal cells (hMSCs)
ROCK inhibitor
MTS assay
Science & Technology
topic Human mesenchymal stem cells
Human multipotent stromal cells (hMSCs)
ROCK inhibitor
MTS assay
Science & Technology
description Y-27632 is a well-known inhibitor of the Rho-associated coiled kinase (ROCK) and has been shown to significantly improve the culture of a variety of multipotent stem cell types. However, the effects of Y-27632 on the expansion of adult human adipose-derived stem cell (hADSC) cultures remain to be established. Here, we aimed to characterize the effects of Y-27632 on the culture of hADSCs. Adult hADSCs were isolated from subjects submitted to elective plastic surgery procedures and cultivated in vitro under optimized conditions. Our results show that the continuous supplementation of hADSC cultures with Y-27632 led to decreased numbers of cells and decreased global metabolic viability of hADSC cultures when compared with control conditions. This effect appeared to be dependent on the continuous presence of the drug and was shown to be concentration-dependent and significant for 10 muM and 20 muM of Y-27632. Moreover, the Y-27632-induced decrease in hADSC numbers was not linked to a block in global cell proliferation, as cell numbers consistently increased from the moment of plating until passaging. In addition, Y-27632 was not able to increase the number of hADSCs present in culture 24 hours after passaging. Taken together, our results suggest that, in contrast to other stem cell types, Y-27632 supplementation is not a suitable strategy to enhance hADSC culture expansion.
publishDate 2015
dc.date.none.fl_str_mv 2015-01-07
2015-01-07T00:00:00Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/1822/34241
url http://hdl.handle.net/1822/34241
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 1178-6957
10.2147/SCCAA.S66597
http://www.dovepress.com
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dc.publisher.none.fl_str_mv Dove Press Ltd
publisher.none.fl_str_mv Dove Press Ltd
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
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collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
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