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KMID : 0848120130380030111
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International Journal of Oral Biology
2013 Volume.38 No. 3 p.111 ~ p.119
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Effects of CoCl₂ on Osteogenic Differentiation of Human Mesenchymal Stem Cells
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Kim Min-Seok
Moon Yeon-Hee
Son Jung-Wan
Moon Jung-Sun
Kang Jee-Hae
Kim Sun-Hun
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Abstract
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Objective. To investigate the effects of the hypoxia induciblefactor-1 (HIF-1) activation?mimicking agent cobalt chloride(CoCl2) on the osteogenic differentiation of human mesenchymalstem cells (hMSCs) and elucidate the underlying molecularmechanisms. Study design. The dose and exposureperiods for CoCl2 in hMSCs were optimized by cell viabilityassays. After confirmation of CoCl2-induced HIF-1α and vascularendothelial growth factor expression in these cells byRT-PCR, the effects of temporary preconditioning with CoCl2on hMSC osteogenic differentiation were evaluated by RTPCRanalysis of osteogenic gene expression, an alkaline phosphatase(ALP) activity assay and by alizarin red S staining.Results. Variable CoCl2 dosages (up to 500 μM) and exposure times(up to 7 days) on hMSC had little effect on hMSC survival.After CoCl2 treatment of hMSCs at 100 μM for 24 or 48hours, followed by culture in osteogenic differentiatingmedia, several osteogenic markers such as Runx-2, osteocalcinand osteopontin, bone sialoprotein mRNA expressionlevel were found to be up-regulated. Moreover, ALP activitywas increased in these treated cells in which an acceleratedosteogenic capacity was also verified by alizarin red Sstaining. Conclusions. The osteogenic differentiation potentialof hMSCs could be preserved and even enhanced byCoCl2 treatment.
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KEYWORD
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MSC, osteogenic differentiation, hypoxia
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