Expression pattern differences between osteoarthritic chondrocytes and mesenchymal stem cells during chondrogenic differentiation P. Bernstein, C. Sticht,

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Expression pattern differences between osteoarthritic chondrocytes and mesenchymal stem cells during chondrogenic differentiation P. Bernstein, C. Sticht, A. Jacobi, C. Liebers, S. Manthey, M. Stiehler Osteoarthritis and Cartilage Volume 18, Issue 12, Pages 1596-1607 (December 2010) DOI: 10.1016/j.joca.2010.09.007 Copyright © 2010 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Data analysis pathway, depicting the three major steps of the analysis process. (1): microarray-screening and filtering through paths A and B by comparing gene expression trends, resulting in gene list 1; (2): hypothesis-driven selection of biological relevant genes, resulting in gene list 2; (3): validation by quantitative RT-PCR. Osteoarthritis and Cartilage 2010 18, 1596-1607DOI: (10.1016/j.joca.2010.09.007) Copyright © 2010 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Immunohistological staining of chondrocyte (A, C, E, G) and MSC pellets (B, D, F, H) reveals sufficient Sox9-expression (A, B) and only limited intranuclear retention in MSC (B). Collagen I is produced on a low level in both cell types (C, D). Collagen II is highly upregulated in chondrocytes (E) as opposed to MSCs (F). Both cell types show an upregulated expression of collagen X (G, H). Sections are representative for larger areas of the specimen. Scale bars=20μm. Osteoarthritis and Cartilage 2010 18, 1596-1607DOI: (10.1016/j.joca.2010.09.007) Copyright © 2010 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Multivariate correlation matrix of genetic expression similarity between chondrocytes and MSCs compared with the monolayer startpoint (day 0) during pellet cultivation until day 14. Line A: Similarity between chondrocytes and MSCs; line B: similarity between dedifferentiated day-0 chondrocytes and pellet-cultured MSCs; line C: similarity between day-0 MSCs and pellet-cultured chondrocytes. Osteoarthritis and Cartilage 2010 18, 1596-1607DOI: (10.1016/j.joca.2010.09.007) Copyright © 2010 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Trend-distribution matrix showing the final difference in gene expression between day 14 and day 3 (dynamics) of selected genes from gene list 1 from chondrocytes and MSCs, cultivated as pellets. A dynamic value>0 represents an increasing expression. Osteoarthritis and Cartilage 2010 18, 1596-1607DOI: (10.1016/j.joca.2010.09.007) Copyright © 2010 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Results of quantitative RT-PCR validation of gene expression differences between chondrocytes and MSC in the first and second week of PC in TGFβ− and TGFβ+ culture conditions (n=6 patients, see material part). P-values of significant differences are indicated above the bars. Error bars show 95% confidence interval. Osteoarthritis and Cartilage 2010 18, 1596-1607DOI: (10.1016/j.joca.2010.09.007) Copyright © 2010 Osteoarthritis Research Society International Terms and Conditions