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. 2014 Jan 13;4:3652. doi: 10.1038/srep03652

Figure 5. SCRG1/BST1 preserves osteogenic differentiation of hMSCs after ex vivo expansion.

Figure 5

(a and b) UE7T-13 cells were cultured in osteogenic differentiation medium (ODM) containing 500 ng/mL (a), or various concentrations of rhSCRG1 (b, 5–500 ng/mL). After 2 weeks, the cells were evaluated for extra cellular matrix mineralization by alizarin red staining (a). Alizarin red was extracted with 10% cetylpyridinium chloride and absorbance was measured at 540 nm (b). (c) UE7T-13 cells were cultured in ODM with 500 ng/mL rhSCRG1 and 1 μM PI3K inhibitor LY294002 (PI3K), MEK inhibitor U0126 (MEK), JNK inhibitor SP600125 (JNK), and FAK inhibitor I (FAK) for 1 week. Total RNAs were extracted and qRT-PCR was performed with tissue non-specific (liver/bone/kidney) alkaline phosphatase (ALPL)-specific oligonucleotide primers. mRNA expression was normalized to GAPDH and results are expressed as fold-increase or decrease relative to the control. (d) Primary cultured hMSCs (passage number #5) were subcultured ten times in the presence (passage number #15, +rhSCRG1) or absence (passage number #15, −rhSCRG1) of 500 ng/mL rhSCRG1. The cells were evaluated for extra cellular matrix mineralization as in (b). In b–d, data are presented as mean ± SD. * p < 0.05 was considered significant.