I. Uzieliene (Vilnius, LT)
State Research Institute Centre for Innovative medicine Regenerative medicinePresenter Of 1 Presentation
P233 - Application of cell sheet technology for human mesenchymal stem cell chondrogenic differentiation
Abstract
Purpose
Human articular cartilage has a weak ability to restore its lesions leading the tissue to highly progressive diseases such as osteoarthritis (OA). Cell-based therapies such as mesenchymal stem cells (MSCs) seem promising therapeutic approach for cartilage engineering and regeneration. Various scaffolds/hydrogels are being used for stimulating MSC chondrogenesis in vitro. However, to date, no effective chondrogenic differentiation technologies using 3D scaffolds exist. Cell sheet technology is a transplantable scaffold-free construct created by stimulating MSC chondrogenesis in vitro, and is promising for further applications. The aim of this study is to stimulate chondrogenic differentiation of bone-marrow MSCs (BMMSCs) using cell sheet technology and to compare it with classical 3D pellet differentiation model. Chondrocytes were used as positive control.
Methods and Materials
BMMSCs were isolated and characterized according to stem cell surface marker expression (flow cytometry) and adipogenic/osteogenic differentiation capacity (Oil-Red/Alizarin). The cells were stimulated to differentiate into chondrogenic lineage in pellet cultures and cell sheets for 21 days. Chondrogenic differentiation was evaluated by the intensity of production of hyaline cartilage extracellular matrix proteins (proteoglycans, collagen type II, COMP) by staining histological sections with safranin-O and immunohistological labeling with antibodies against collagen type II, as well as using ELISA and RT-PCR.
Results
According to histological sample staining, BMMSC differentiate more effectively in cell sheet model, while chondrocyte differentiation was more effective in pellet system. Gene expression analysis showed that during chondrogenic differentiation in cell sheets, cells actively synthesized early transcription factor of chondrogenesis SOX9, and aggrecan. Moreover, cartilage oligomeric matrix protein (COMP) secretion was analysed during chondrogenic differentiation (ELISA). Higher COMP concentration was observed in pellet model supernatants, as compared to cell sheets, which is associated with higher extracellular matrix degradation in pellet system.
Conclusion
In conclusion, cell sheet technology is a promising chondrogenesis stimulating method for BMMSCs and potential application for further cartilage tissue regeneration.
Presenter Of 1 Presentation
P233 - Application of cell sheet technology for human mesenchymal stem cell chondrogenic differentiation
Abstract
Purpose
Human articular cartilage has a weak ability to restore its lesions leading the tissue to highly progressive diseases such as osteoarthritis (OA). Cell-based therapies such as mesenchymal stem cells (MSCs) seem promising therapeutic approach for cartilage engineering and regeneration. Various scaffolds/hydrogels are being used for stimulating MSC chondrogenesis in vitro. However, to date, no effective chondrogenic differentiation technologies using 3D scaffolds exist. Cell sheet technology is a transplantable scaffold-free construct created by stimulating MSC chondrogenesis in vitro, and is promising for further applications. The aim of this study is to stimulate chondrogenic differentiation of bone-marrow MSCs (BMMSCs) using cell sheet technology and to compare it with classical 3D pellet differentiation model. Chondrocytes were used as positive control.
Methods and Materials
BMMSCs were isolated and characterized according to stem cell surface marker expression (flow cytometry) and adipogenic/osteogenic differentiation capacity (Oil-Red/Alizarin). The cells were stimulated to differentiate into chondrogenic lineage in pellet cultures and cell sheets for 21 days. Chondrogenic differentiation was evaluated by the intensity of production of hyaline cartilage extracellular matrix proteins (proteoglycans, collagen type II, COMP) by staining histological sections with safranin-O and immunohistological labeling with antibodies against collagen type II, as well as using ELISA and RT-PCR.
Results
According to histological sample staining, BMMSC differentiate more effectively in cell sheet model, while chondrocyte differentiation was more effective in pellet system. Gene expression analysis showed that during chondrogenic differentiation in cell sheets, cells actively synthesized early transcription factor of chondrogenesis SOX9, and aggrecan. Moreover, cartilage oligomeric matrix protein (COMP) secretion was analysed during chondrogenic differentiation (ELISA). Higher COMP concentration was observed in pellet model supernatants, as compared to cell sheets, which is associated with higher extracellular matrix degradation in pellet system.
Conclusion
In conclusion, cell sheet technology is a promising chondrogenesis stimulating method for BMMSCs and potential application for further cartilage tissue regeneration.