Thickness-wise growth technique for human articular chondrocytes to fabricate three-dimensional cartilage grafts
| Autor: | Tatsuya Shimizu, Tetsutaro Kikuchi |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
GDF Growth and differentiation factor Biomedical Engineering FGF-2 Fibroblast growth factor 2 Regenerative medicine Chondrocyte Biomaterials 03 medical and health sciences 0302 clinical medicine Tissue engineering MIA Melanoma inhibitory activity medicine Cartilaginous Tissue TGF-β1 Transforming growth factor β1 lcsh:QH573-671 Fibroblast BMP Bone morphogenetic protein lcsh:R5-920 biology Chemistry OCT Optical coherence tomography lcsh:Cytology Cartilage Melanoma inhibitory activity Transforming growth factor beta Cell biology Three-dimensional culture 030104 developmental biology medicine.anatomical_structure biology.protein Original Article lcsh:Medicine (General) 030217 neurology & neurosurgery Developmental Biology |
| Zdroj: | Regenerative Therapy, Vol 14, Iss, Pp 119-127 (2020) Regenerative Therapy |
| ISSN: | 2352-3204 |
| Popis: | Introduction Cutting the cost of manufacturing is important for extending the use of tissue-engineered therapeutic products. The present study aimed to develop a simple method for fabrication of cartilaginous tissues for regenerative therapy, utilizing the phenomenon where human articular chondrocytes grow thickness-wise and spontaneously form three-dimensionally thick tissues. Methods Normal human articular chondrocytes (NHACs) were cultured with varying concentrations of transforming growth factor beta 1 (TGF-β1) and/or fibroblast growth factor-2 (FGF-2) to optimize the culture condition for thickness-wise growth of chondrocytes. Next, the tissues grown in the optimal condition were subjected to re-differentiation culture in attached and detached states to assess differentiation capacity by evaluating secreted factors, histological analysis, and a gene expression assay. Results NHACs grew thickness-wise efficiently in the presence of 1 ng/mL TGF-β1 and 10 ng/mL FGF-2. After two weeks of culture, NHACs grew with 11-fold higher thickness and 16-fold higher cell number compared to cells which were neither treated with TGF-β1 nor with FGF-2. These thickness-wise-grown chondrocytes could be re-differentiated by a differentiation medium according to the increase in melanoma inhibitory activity (MIA) and positive safranin-O staining. Interestingly, the cartilaginous gene expression was considerably different between the attached and detached conditions even in the same culture medium, indicating the necessity of detachment and shrinkage to achieve further differentiation. Conclusions Spontaneous thickness-wise growth might provide a simple tissue-engineering method for manufacturing cartilaginous 3D tissues. Graphical abstract Image 1 Highlights • Chondrocytes grow thickness-wise efficiently in the presence of FGF-2 and TGF-β1 to medium. • Differentiation medium leads to MIA secretion and Safranin-O staining. • Detachment from culture surface promoted differentiation of chondrocytes. |
| Databáze: | OpenAIRE |
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