Beneficial Effects of Coculturing Synovial Derived Mesenchymal Stem Cells with Meniscus Fibrochondrocytes Are Mediated by Fibroblast Growth Factor 1: Increased Proliferation and Collagen Synthesis

Figure 4

Increased collagen synthesis of me-CH by conditioned medium of SSCs is mediated by FGF1. (a) ELISA was performed to measure collagen type I (a) or collagen type II (b) in me-CH cultured in SF medium (serum free medium plus 5 μg/mL normal goat IgG), Con medium (conditioned medium plus 5 μg/mL normal goat IgG), or Con medium + anti-FGF1 (conditioned medium plus 5 μg/mL of goat antibody against FGF1). Collagen amount was normalized to total DNA in pellets (). values were calculated with one-way ANOVA followed by Dunnett’s test. (b) Quantitative GAG assay (). Error bar reflects standard deviation. NS: nonsignificance. (c) Expressions of three collagens (1a1, 2a1, and 9a1) were measured by qPCR in me-CH pellets cultured in the same three conditions. GAPDH was used for normalization. me-CH in SF medium was chosen as reference. Number in coculture represents the relative expression level of corresponding gene compared to me-CH. Three donor pairs were analyzed. (d) Total protein was extracted from me-CH cultured in the same three conditions for 1 hour in 2D environment. Western blot was then performed to detect the phosphorylation of MEK1/2 and ERK1/2. Total MEK1/2 and ERK1/2 were shown as controls. , when comparing Con medium group with SF medium group. , when comparing Con medium + anti-FGF1 group with Con medium group. , when comparing Con medium group with SF medium group.

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