Poly-L-lysine Prevents Senescence and Augments Growth in Culturing Mesenchymal Stem Cells Ex Vivo
Mesenchymal stem cells (MSCs) possess great therapeutic potential. Efficient in vitro expansion of MSCs is however necessary for their clinical application. The extracellular matrix (ECM) provides structural and biochemical support to the surrounding cells, and it has been used as a coating substrat...
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Online Access: | http://dx.doi.org/10.1155/2016/8196078 |
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doaj-c0acc3b1d43f4ce983806309f4899de82020-11-24T23:48:48ZengHindawi LimitedBioMed Research International2314-61332314-61412016-01-01201610.1155/2016/81960788196078Poly-L-lysine Prevents Senescence and Augments Growth in Culturing Mesenchymal Stem Cells Ex VivoJune Seok Heo0Hyun Ok Kim1Seung Yong Song2Dae Hyun Lew3Youjeong Choi4Sinyoung Kim5Cell Therapy Centre, Severance Hospital, Seoul 03722, Republic of KoreaCell Therapy Centre, Severance Hospital, Seoul 03722, Republic of KoreaDepartment of Plastic and Reconstructive Surgery, Yonsei University College of Medicine, Seoul 03722, Republic of KoreaDepartment of Plastic and Reconstructive Surgery, Yonsei University College of Medicine, Seoul 03722, Republic of KoreaCell Therapy Centre, Severance Hospital, Seoul 03722, Republic of KoreaDepartment of Laboratory Medicine, Yonsei University College of Medicine, Seoul 03722, Republic of KoreaMesenchymal stem cells (MSCs) possess great therapeutic potential. Efficient in vitro expansion of MSCs is however necessary for their clinical application. The extracellular matrix (ECM) provides structural and biochemical support to the surrounding cells, and it has been used as a coating substrate for cell culture. In this study, we have aimed to improve the functionality and stemness of MSCs during culture using poly-L-lysine (PLL). Functionality of MSCs was analysed by cell cycle analysis, differentiation assay, β-galactosidase staining, and RT-PCR. Furthermore, we assessed the global gene expression profile of MSCs on uncoated and PLL-coated plates. MSCs on PLL-coated plates exhibited a faster growth rate with increased S-phase and upregulated expression of the stemness markers. In addition, their osteogenic differentiation potential was increased, and genes involved in cell adhesion, FGF-2 signalling, cell cycle, stemness, cell differentiation, and cell proliferation were upregulated, compared to that of the MSCs cultured on uncoated plates. We also confirmed that MSCs on uncoated plates expressed higher β-galactosidase than the MSCs on PLL-coated plates. We demonstrate that PLL provides favourable microenvironment for MSC culture by reversing the replicative senescence. This method will significantly contribute to effective preparation of MSCs for cellular therapy.http://dx.doi.org/10.1155/2016/8196078 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
June Seok Heo Hyun Ok Kim Seung Yong Song Dae Hyun Lew Youjeong Choi Sinyoung Kim |
spellingShingle |
June Seok Heo Hyun Ok Kim Seung Yong Song Dae Hyun Lew Youjeong Choi Sinyoung Kim Poly-L-lysine Prevents Senescence and Augments Growth in Culturing Mesenchymal Stem Cells Ex Vivo BioMed Research International |
author_facet |
June Seok Heo Hyun Ok Kim Seung Yong Song Dae Hyun Lew Youjeong Choi Sinyoung Kim |
author_sort |
June Seok Heo |
title |
Poly-L-lysine Prevents Senescence and Augments Growth in Culturing Mesenchymal Stem Cells Ex Vivo |
title_short |
Poly-L-lysine Prevents Senescence and Augments Growth in Culturing Mesenchymal Stem Cells Ex Vivo |
title_full |
Poly-L-lysine Prevents Senescence and Augments Growth in Culturing Mesenchymal Stem Cells Ex Vivo |
title_fullStr |
Poly-L-lysine Prevents Senescence and Augments Growth in Culturing Mesenchymal Stem Cells Ex Vivo |
title_full_unstemmed |
Poly-L-lysine Prevents Senescence and Augments Growth in Culturing Mesenchymal Stem Cells Ex Vivo |
title_sort |
poly-l-lysine prevents senescence and augments growth in culturing mesenchymal stem cells ex vivo |
publisher |
Hindawi Limited |
series |
BioMed Research International |
issn |
2314-6133 2314-6141 |
publishDate |
2016-01-01 |
description |
Mesenchymal stem cells (MSCs) possess great therapeutic potential. Efficient in vitro expansion of MSCs is however necessary for their clinical application. The extracellular matrix (ECM) provides structural and biochemical support to the surrounding cells, and it has been used as a coating substrate for cell culture. In this study, we have aimed to improve the functionality and stemness of MSCs during culture using poly-L-lysine (PLL). Functionality of MSCs was analysed by cell cycle analysis, differentiation assay, β-galactosidase staining, and RT-PCR. Furthermore, we assessed the global gene expression profile of MSCs on uncoated and PLL-coated plates. MSCs on PLL-coated plates exhibited a faster growth rate with increased S-phase and upregulated expression of the stemness markers. In addition, their osteogenic differentiation potential was increased, and genes involved in cell adhesion, FGF-2 signalling, cell cycle, stemness, cell differentiation, and cell proliferation were upregulated, compared to that of the MSCs cultured on uncoated plates. We also confirmed that MSCs on uncoated plates expressed higher β-galactosidase than the MSCs on PLL-coated plates. We demonstrate that PLL provides favourable microenvironment for MSC culture by reversing the replicative senescence. This method will significantly contribute to effective preparation of MSCs for cellular therapy. |
url |
http://dx.doi.org/10.1155/2016/8196078 |
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