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

Nuno Jorge Lamas,1–3 Sofia C Serra,1,2 António J Salgado,1,2 Nuno Sousa1,2 1Life and Health Sciences Research Institute (ICVS), School of Health Sciences (ECS), University of Minho, Braga, Portugal; 2ICVS/3B’s-PT Government Associate Laboratory, Braga/Guimar&atild...

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Autores principales:	Lamas NJ, Serra SC, Salgado AJ, Sousa N
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2015
Materias:	Cytology QH573-671
Acceso en línea:	https://doaj.org/article/7f6b6074b27c487da1ba2c160c253742
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spelling	oai:doaj.org-article:7f6b6074b27c487da1ba2c160c2537422021-12-02T07:08:33ZFailure of Y-27632 to improve the culture of adult human adipose-derived stem cells1178-6957https://doaj.org/article/7f6b6074b27c487da1ba2c160c2537422015-01-01T00:00:00Zhttp://www.dovepress.com/failure-of-y-27632-to-improve-the-culture-of-adult-human-adipose-deriv-peer-reviewed-article-SCCAAhttps://doaj.org/toc/1178-6957 Nuno Jorge Lamas,1–3 Sofia C Serra,1,2 António J Salgado,1,2 Nuno Sousa1,2 1Life and Health Sciences Research Institute (ICVS), School of Health Sciences (ECS), University of Minho, Braga, Portugal; 2ICVS/3B’s-PT Government Associate Laboratory, Braga/Guimarães, Portugal; 3Clinical Pathology Department, Centro Hospitalar do Alto Ave (CHAA), EPE, Guimarães, Portugal Abstract: 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 µM and 20 µM 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. Keywords: human mesenchymal stem cells, human multipotent stromal cells (hMSCs), ROCK inhibitor, MTS assayLamas NJSerra SCSalgado AJSousa NDove Medical PressarticleCytologyQH573-671ENStem Cells and Cloning: Advances and Applications, Vol 2015, Iss default, Pp 15-26 (2015)
institution	DOAJ
collection	DOAJ
language	EN
topic	Cytology QH573-671
spellingShingle	Cytology QH573-671 Lamas NJ Serra SC Salgado AJ Sousa N Failure of Y-27632 to improve the culture of adult human adipose-derived stem cells
description	Nuno Jorge Lamas,1–3 Sofia C Serra,1,2 António J Salgado,1,2 Nuno Sousa1,2 1Life and Health Sciences Research Institute (ICVS), School of Health Sciences (ECS), University of Minho, Braga, Portugal; 2ICVS/3B’s-PT Government Associate Laboratory, Braga/Guimarães, Portugal; 3Clinical Pathology Department, Centro Hospitalar do Alto Ave (CHAA), EPE, Guimarães, Portugal Abstract: 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 µM and 20 µM 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. Keywords: human mesenchymal stem cells, human multipotent stromal cells (hMSCs), ROCK inhibitor, MTS assay
format	article
author	Lamas NJ Serra SC Salgado AJ Sousa N
author_facet	Lamas NJ Serra SC Salgado AJ Sousa N
author_sort	Lamas NJ
title	Failure of Y-27632 to improve the culture of adult human adipose-derived stem cells
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
publisher	Dove Medical Press
publishDate	2015
url	https://doaj.org/article/7f6b6074b27c487da1ba2c160c253742
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Failure of Y-27632 to improve the culture of adult human adipose-derived stem cells

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