Artikel
Optimizing the chondrogenic differentiation of adipose tissue derived stem cells (ADSC) in collagen gels
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Veröffentlicht: | 3. August 2011 |
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Gliederung
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Introduction: Adipose tissue derived stem cells (ADSC) can be trans-differentiated to matrix producing chondrocytes. The primary goal is the development of methods for surgical tissue augmentation.
Methods: After isolation of ADSC from human liposuction material and subsequent cell growth to the passages 2 to 4, the cartilage cell formation in collagen type I-hydrogels was induced. This was performed with 3 different cell densities (1x105, 5x105 und 1x106 per ml) by addition of TGFß-3 over a maximum of 28 days. Chondrogenesis was verified with alcian blue staining and immunofluorescence with the cartilage specific antibodies anti-collagen type II, anti-aggrecan and anti-Sox9. The content of cartilage specific glycosaminoglycans, the content of total collagen and the expression of cartilage specific genetic markers by means of real-time PCR was determined. Native human cartilage served as positive control and uninduced ADSC as negative control for the detection methods.
Results: RT-PCR showed an increase of the expression of the analyzed cartilage-specific genetic markers from day 14 on after addition of growth factor TGFß-3. Positive alcian blue staining showed the production of proteoglycans from day 14 on.
Conclusions: The in vitro differentiation of human ADSC to matrix producing chondrocytes in an injectable supporting material was achieved. A cartilaginous matrix formation in the collagen gels by cartilage specific matrix proteins could be shown. The comparison of the results of the different cell densities in collagen gels with the native cartilage was performed in order to determine the optimal cell density and induction period of the ADSC. Future in vivo applications will be tested in the animal model.