Functional Diversity of Fibroblast Growth Factors in Bone Formation

Figure 2

Expression profiling of Fgf genes in rat calvaria cell cultures. (a) Outline of osteoblast development. Rat calvaria cells from 21-day-old fetal rats [9] were plated at 3,000 cells per cm² and grown in αMEM supplemented with 10% fetal calf serum plus 50 μg/mL ascorbic acid. Cells proliferate, reach confluence at day 6, and subsequently initiate osteoid-like nodule formation. To determine matrix mineralization, 10 mM β-glycerophosphate (βGP) is added to cultures for 2 days before culture termination. (b) Distinct gene expression patterns of Fgfs during osteoblast development. Total RNA was routinely prepared as indicated time points, and cDNA synthesis and quantitative real-time RT PCR (qPCR) were performed using standard protocols. Ribosomal protein L32 was used as internal control. Data represent means ± S.D. . Statistical significance of differences was analyzed with one-way or two-way analysis of variance (ANOVA) with repeated measures, followed by Tukey’s multiple comparison test. and versus day 3.

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