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The changing integrin expression and a role for integrin β8 in the chondrogenic differentiation of mesenchymal stem cells.

LaPointe VL, Verpoorte A, Stevens MM - PLoS ONE (2013)

Bottom Line: We found that transcript expression of most subunits was conserved across the chondrogenesis models, but was significantly affected by the time-course of differentiation.In conclusion, we performed a systematic study of the temporal changes of integrin expression during chondrogenic differentiation in multiple chondrogenesis models, and revealed a role for integrin β8 in chondrogenesis.This work enhances our understanding of the changing adhesion requirements of hMSCs during chondrogenic differentiation and underlines the importance of integrins in establishing a cartilage phenotype.

View Article: PubMed Central - PubMed

Affiliation: Departments of Materials and Bioengineering, and the Institute of Biomedical Engineering, Imperial College London, London, United Kingdom.

ABSTRACT
Many cartilage tissue engineering approaches aim to differentiate human mesenchymal stem cells (hMSCs) into chondrocytes and develop cartilage in vitro by targeting cell-matrix interactions. We sought to better inform the design of cartilage tissue engineering scaffolds by understanding how integrin expression changes during chondrogenic differentiation. In three models of in vitro chondrogenesis, we studied the temporal change of cartilage phenotype markers and integrin subunits during the differentiation of hMSCs. We found that transcript expression of most subunits was conserved across the chondrogenesis models, but was significantly affected by the time-course of differentiation. In particular, ITGB8 was up-regulated and its importance in chondrogenesis was further established by a knockdown of integrin β8, which resulted in a non-hyaline cartilage phenotype, with no COL2A1 expression detected. In conclusion, we performed a systematic study of the temporal changes of integrin expression during chondrogenic differentiation in multiple chondrogenesis models, and revealed a role for integrin β8 in chondrogenesis. This work enhances our understanding of the changing adhesion requirements of hMSCs during chondrogenic differentiation and underlines the importance of integrins in establishing a cartilage phenotype.

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To study the changing expression of integrins during chondrogenic differentiation, human mesenchymal stem cells (hMSCs) were cultured in three different chondrogenesis models: pellet culture, micromass, and type II collagen hydrogels under two different conditions: growth medium and chondrogenic medium.
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pone-0082035-g001: To study the changing expression of integrins during chondrogenic differentiation, human mesenchymal stem cells (hMSCs) were cultured in three different chondrogenesis models: pellet culture, micromass, and type II collagen hydrogels under two different conditions: growth medium and chondrogenic medium.

Mentions: Before determining the changing integrin expression during chondrogenesis, we sought an appropriate in vitro model that resulted in a high expression of chondrogenic markers and the establishment of a hyaline cartilage-like extracellular matrix (ECM). We therefore used qPCR to characterise three different chondrogenesis models (Figure 1; pellet culture, micromass culture, and a type II collagen hydrogel) for the expression of COL1A1 (type I collagen), COL2A1 (type II collagen), COL6A1 (type VI collagen), COL10A1 (type X collagen), ACAN (aggrecan), HSPG2 (perlecan), RUNX2, and SOX9 (Figures 2-3), and also used histology to view an ECM rich in collagen and sulphated glycosaminoglycans (Figure S1). In all experiments, results are reported as statistically significant when p < 0.05.


The changing integrin expression and a role for integrin β8 in the chondrogenic differentiation of mesenchymal stem cells.

LaPointe VL, Verpoorte A, Stevens MM - PLoS ONE (2013)

To study the changing expression of integrins during chondrogenic differentiation, human mesenchymal stem cells (hMSCs) were cultured in three different chondrogenesis models: pellet culture, micromass, and type II collagen hydrogels under two different conditions: growth medium and chondrogenic medium.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3842320&req=5

pone-0082035-g001: To study the changing expression of integrins during chondrogenic differentiation, human mesenchymal stem cells (hMSCs) were cultured in three different chondrogenesis models: pellet culture, micromass, and type II collagen hydrogels under two different conditions: growth medium and chondrogenic medium.
Mentions: Before determining the changing integrin expression during chondrogenesis, we sought an appropriate in vitro model that resulted in a high expression of chondrogenic markers and the establishment of a hyaline cartilage-like extracellular matrix (ECM). We therefore used qPCR to characterise three different chondrogenesis models (Figure 1; pellet culture, micromass culture, and a type II collagen hydrogel) for the expression of COL1A1 (type I collagen), COL2A1 (type II collagen), COL6A1 (type VI collagen), COL10A1 (type X collagen), ACAN (aggrecan), HSPG2 (perlecan), RUNX2, and SOX9 (Figures 2-3), and also used histology to view an ECM rich in collagen and sulphated glycosaminoglycans (Figure S1). In all experiments, results are reported as statistically significant when p < 0.05.

Bottom Line: We found that transcript expression of most subunits was conserved across the chondrogenesis models, but was significantly affected by the time-course of differentiation.In conclusion, we performed a systematic study of the temporal changes of integrin expression during chondrogenic differentiation in multiple chondrogenesis models, and revealed a role for integrin β8 in chondrogenesis.This work enhances our understanding of the changing adhesion requirements of hMSCs during chondrogenic differentiation and underlines the importance of integrins in establishing a cartilage phenotype.

View Article: PubMed Central - PubMed

Affiliation: Departments of Materials and Bioengineering, and the Institute of Biomedical Engineering, Imperial College London, London, United Kingdom.

ABSTRACT
Many cartilage tissue engineering approaches aim to differentiate human mesenchymal stem cells (hMSCs) into chondrocytes and develop cartilage in vitro by targeting cell-matrix interactions. We sought to better inform the design of cartilage tissue engineering scaffolds by understanding how integrin expression changes during chondrogenic differentiation. In three models of in vitro chondrogenesis, we studied the temporal change of cartilage phenotype markers and integrin subunits during the differentiation of hMSCs. We found that transcript expression of most subunits was conserved across the chondrogenesis models, but was significantly affected by the time-course of differentiation. In particular, ITGB8 was up-regulated and its importance in chondrogenesis was further established by a knockdown of integrin β8, which resulted in a non-hyaline cartilage phenotype, with no COL2A1 expression detected. In conclusion, we performed a systematic study of the temporal changes of integrin expression during chondrogenic differentiation in multiple chondrogenesis models, and revealed a role for integrin β8 in chondrogenesis. This work enhances our understanding of the changing adhesion requirements of hMSCs during chondrogenic differentiation and underlines the importance of integrins in establishing a cartilage phenotype.

Show MeSH
Related in: MedlinePlus