Artikel
Chondrogenic differentiation of mesenchymal stem cells with combinations of different hydrogels and growth factors
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Veröffentlicht: | 30. Juli 2013 |
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Gliederung
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Introduction: Chondrogenesis of adipose tissue derived stem cells (ASCs) can be induced using various growth factors and scaffold materials. The aim of the present study was to examine and evaluate the chondrogenic differentiation of ASCs under various growth factor conditions in hydrogels.
Methods: ASCs were isolated from human liposuction material of 4 patients and cultured for 2 to 4 passages. Afterwards, cells were seeded with a density of 1 million / ml in 1:1 hydrogel combinations of collagen type I, agarose, fibrin, and hyaluronic acid. Chondrogenesis was induced by adding combinations of the growth factors TGF-ß3, BMP-6 and IGF-I over a 14 day period. The confirmation of chondrogenesis was performed with the histological stains Alcian Blue, Safranin O and Picrosirius Red. In addition, the content of cartilage-specific glycosaminoglycans and total collagen was determined. The expression of cartilage-specific marker genes was analyzed using Real-Time PCR.
Results: The PCR showed the expression of cartilage-specific marker genes after cultivation of the cells with above mentioned growth factors. Positive histological staining and biochemical assays confirmed the production of cartilage specific matrix proteins. This was pronounced in the agarose free hydrogels that have been cultivated with TGF-ß3 plus BMP-6 and TGF-ß3 plus IGF-I, respectively.
Conclusion: ASCs can be differentiated into matrix-producing chondrocytes in hydrogels in vitro. The evidence of cartilage-specific proteins in the hydrogels suggests a cartilage-like matrix formation. The evaluation of the different hydrogel combinations and growth factors turned out that agarose free hydrogels are optimal scaffolds which should be tested in an animal model in vivo.