Fig. 4.

S1P increases fHASC differentiation towards specific cell lineages. fHASCs were pre-treatment with 1 μM S1P for 24 h in basal stem-cell culture medium then cultured in specific differentiation media for adipogenic, osteogenic or endothelial cell lineage differentiation. A) mRNA levels of specific adipogenic markers. Data (means ± S.D) are presented as normalized transcript expressions in the samples relative to normalized transcript expression in control medium in one experiment representative of two. ***P < 0.001 by ANOVA followed by Bonferroni multiple comparison test. B) representative pictures of fHASC differentiation into adipocytes monitored with Sudan III staining for lipid droplets and nuclear staining with Mayer's hematoxylin, scale bar = 100 μm. Sudan III positive stained areas (means ± S.D.) were determined. C) mRNA levels of specific osteogenic markers. Data (means ± S.D.) are showed as normalized transcript expressions in the samples relative to normalized transcript expression in samples cultured in control medium. One experiment representative of two. **P < 0.01 by ANOVA followed by Bonferroni multiple comparison test. D) Representative pictures of fHASC differentiation in osteocytes monitored with Von Kossa staining of calcium deposition in the extracellular matrix in terminal osteoblast differentiation and nuclear staining with Mayer's hematoxylin, scale bar = 100 μm. Von Kossa positive stained areas (means ± S.D) were determined. E) mRNA levels of specific endothelial markers. Data (means ± S.D) presented as normalized transcript expressions in the samples relative to normalized those in samples cultured in control medium. One experiment representative of two. ***P < 0.001 by Student's t-test. F) Expression of endothelial cell differentiation markers by FACS, in one representative experiment of three.