Gene expression profiling of chondrogenic differentiation by dexamethasone-conjugated polyethyleneimine with SOX trio genes in stem cells

Abstract Background During differentiation of stem cells, it is recognized that molecular mechanisms of transcription factors manage stem cells towards the intended lineage. In this study, using microarray-based technology, gene expression profiling was examined during the process of chondrogenic di...

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Main Authors: Se Won Yi, Hye Jin Kim, Hyun Jyung Oh, Heejun Shin, Jung Sun Lee, Ji Sun Park, Keun-Hong Park
Format: Article
Language:English
Published: BMC 2018-12-01
Series:Stem Cell Research & Therapy
Subjects:
Online Access:http://link.springer.com/article/10.1186/s13287-018-0998-7
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author Se Won Yi
Hye Jin Kim
Hyun Jyung Oh
Heejun Shin
Jung Sun Lee
Ji Sun Park
Keun-Hong Park
author_facet Se Won Yi
Hye Jin Kim
Hyun Jyung Oh
Heejun Shin
Jung Sun Lee
Ji Sun Park
Keun-Hong Park
author_sort Se Won Yi
collection DOAJ
description Abstract Background During differentiation of stem cells, it is recognized that molecular mechanisms of transcription factors manage stem cells towards the intended lineage. In this study, using microarray-based technology, gene expression profiling was examined during the process of chondrogenic differentiation of human mesenchymal stem cells (hMSCs). To induce chondrogenic differentiation of hMSCs, the cationic polymer polyethyleneimine (PEI) was coupled with the synthetic glucocorticoid dexamethasone (DEX). DEX/PEI could be polyplexed with anionic plasmid DNAs (pDNAs) harboring the chondrogenesis-inducing factors SOX5, SOX6, and SOX9. These are named differentiation-inducing nanoparticles (DI-NPs). Methods A DI-NP system for inducing chondrogenic differentiation was designed and characterized by dynamic light scattering and scanning electron microscopy (SEM). Chondrogenic induction of hMSCs was evaluated using various tools such as reverse-transcription polymerase chain reaction (RT-PCR), Western blotting, confocal fluorescent microscopy, and immunohistochemistry analysis. The gene expression profiling of DI-NP-treated hMSCs was performed by microarray analysis. Results The hMSCs were more efficiently transfected with pDNAs using DI-NPs than using PEI. Moreover, microarray analysis demonstrated the gene expression profiling of hMSCs transfected with DI-NPs. Chondrogenic factors including SOX9, collagen type II (COLII), Aggrecan, and cartilage oligometric matrix protein (COMP) were upregulated while osteogenic factors including collagen type I (COLI) was downregulated. Chondrogenesis-induced hMSCs were better differentiated as assessed by RT-PCR, Western blotting analyses, and immunohistochemistry. Conclusion DI-NPs are good gene delivery carriers and induce chondrogenic differentiation of hMSCs. Additionally, comprehensive examination of the gene expression was attempted to identify specific genes related to differentiation by microarray analysis. Graphical abstract
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spelling doaj.art-e496d72f3c1441828f530a57d18891b82022-12-22T02:02:13ZengBMCStem Cell Research & Therapy1757-65122018-12-019111310.1186/s13287-018-0998-7Gene expression profiling of chondrogenic differentiation by dexamethasone-conjugated polyethyleneimine with SOX trio genes in stem cellsSe Won Yi0Hye Jin Kim1Hyun Jyung Oh2Heejun Shin3Jung Sun Lee4Ji Sun Park5Keun-Hong Park6Department of Nano-regenerative Medical Engineering, College of Life Science, CHA UniversityDepartment of Nano-regenerative Medical Engineering, College of Life Science, CHA UniversityDepartment of Nano-regenerative Medical Engineering, College of Life Science, CHA UniversityDepartment of Biotechnology, Catholic University 43-1Department of Nano-regenerative Medical Engineering, College of Life Science, CHA UniversityDepartment of Nano-regenerative Medical Engineering, College of Life Science, CHA UniversityDepartment of Nano-regenerative Medical Engineering, College of Life Science, CHA UniversityAbstract Background During differentiation of stem cells, it is recognized that molecular mechanisms of transcription factors manage stem cells towards the intended lineage. In this study, using microarray-based technology, gene expression profiling was examined during the process of chondrogenic differentiation of human mesenchymal stem cells (hMSCs). To induce chondrogenic differentiation of hMSCs, the cationic polymer polyethyleneimine (PEI) was coupled with the synthetic glucocorticoid dexamethasone (DEX). DEX/PEI could be polyplexed with anionic plasmid DNAs (pDNAs) harboring the chondrogenesis-inducing factors SOX5, SOX6, and SOX9. These are named differentiation-inducing nanoparticles (DI-NPs). Methods A DI-NP system for inducing chondrogenic differentiation was designed and characterized by dynamic light scattering and scanning electron microscopy (SEM). Chondrogenic induction of hMSCs was evaluated using various tools such as reverse-transcription polymerase chain reaction (RT-PCR), Western blotting, confocal fluorescent microscopy, and immunohistochemistry analysis. The gene expression profiling of DI-NP-treated hMSCs was performed by microarray analysis. Results The hMSCs were more efficiently transfected with pDNAs using DI-NPs than using PEI. Moreover, microarray analysis demonstrated the gene expression profiling of hMSCs transfected with DI-NPs. Chondrogenic factors including SOX9, collagen type II (COLII), Aggrecan, and cartilage oligometric matrix protein (COMP) were upregulated while osteogenic factors including collagen type I (COLI) was downregulated. Chondrogenesis-induced hMSCs were better differentiated as assessed by RT-PCR, Western blotting analyses, and immunohistochemistry. Conclusion DI-NPs are good gene delivery carriers and induce chondrogenic differentiation of hMSCs. Additionally, comprehensive examination of the gene expression was attempted to identify specific genes related to differentiation by microarray analysis. Graphical abstracthttp://link.springer.com/article/10.1186/s13287-018-0998-7hMSCDexamethasonePEIDI-NPSOX triomRNA profiling
spellingShingle Se Won Yi
Hye Jin Kim
Hyun Jyung Oh
Heejun Shin
Jung Sun Lee
Ji Sun Park
Keun-Hong Park
Gene expression profiling of chondrogenic differentiation by dexamethasone-conjugated polyethyleneimine with SOX trio genes in stem cells
Stem Cell Research & Therapy
hMSC
Dexamethasone
PEI
DI-NP
SOX trio
mRNA profiling
title Gene expression profiling of chondrogenic differentiation by dexamethasone-conjugated polyethyleneimine with SOX trio genes in stem cells
title_full Gene expression profiling of chondrogenic differentiation by dexamethasone-conjugated polyethyleneimine with SOX trio genes in stem cells
title_fullStr Gene expression profiling of chondrogenic differentiation by dexamethasone-conjugated polyethyleneimine with SOX trio genes in stem cells
title_full_unstemmed Gene expression profiling of chondrogenic differentiation by dexamethasone-conjugated polyethyleneimine with SOX trio genes in stem cells
title_short Gene expression profiling of chondrogenic differentiation by dexamethasone-conjugated polyethyleneimine with SOX trio genes in stem cells
title_sort gene expression profiling of chondrogenic differentiation by dexamethasone conjugated polyethyleneimine with sox trio genes in stem cells
topic hMSC
Dexamethasone
PEI
DI-NP
SOX trio
mRNA profiling
url http://link.springer.com/article/10.1186/s13287-018-0998-7
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