Article
A coculture model: Paracrine interaction between cartilage and mesenchymal stem cells
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Published: | October 19, 2004 |
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Outline
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Introduction
Damaged articular cartilage has limited capacity to regenerate mainly because of lack of chondroprogenitor cells. Pluripotent mesenchymal stem cell's (MSCs) potential to differentiate into tissues of mesenchymal lineage including cartilage makes them a good candidate for cartilage repair studies. The purpose of this study is to determine chondrogenic potential of MSCs under paracrine influence of cartilage. A coculture system between cartilage and MSCs embedded in alginate beads is being established.
Methods
A coculture system is set up in 6 well culture plates holding inserts of 1 micron pore size. Cartilage chips are kept in the bottom of well and MSCs suspended in alginate beads are kept in inserts. Cell culture is carried out in serum free chondrogenic medium containing tgfß-3 for four weeks. To ascertain validity of the model, RNA is extracted from the cells at four time points and the molecular response is determined with RT-PCR.
Results
Preliminary results show an increase in Collagen II and Aggrecan gene expression, which is a satisfactory indication of extracellular matrix (ECM) formation. Other related genes as COMP and Collagen IX are also being tested. To further prove the soundness of the system immunohistochemical studies are being carried out. Micro array studies, to identify up or down regulated genes, are also to be carried out.
Conclusions
This system provides mechanical support and a suitable micro-environment for the newly formed ECM under the influence of Chondrogenic medium. With this model the effect of cell signalling from differentiated chondrocytes on MSCs could be studied.