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KMID : 1007020040020010037
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Korean Soceity of Osteroporosis
2004 Volume.2 No. 1 p.37 ~ p.47
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Calcium concentration Effect on Mesenchymal Stem Cell Osteoinduction Using Dexamethasone
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Jang Hyon-Seok
Rim Jae-Suk
Lee Eui-Seok
Yun Jung-Ju
Kim Ho-Sung
Kim Tak
Park Jung-Ho
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Abstract
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Objectives: hMSCs can differentiate along the osteoblastic pathway with osteogenic supplements that include ascorbic acid, β-glycerophosphate and dexamethasone. Alternatives to dexamethasone have been sought to preclude its use in potential differentiation schemes for clinical use. The purpose of this study is to identify relatively simple modifications of the in vitro culture environment that enhances the differentiation of hMSCs using Dexamethasone. As a first approach to this problem, the cell culture media calcium concentration effects on osteoinduction of cultured hMSCs were examined.
Materials and Methods: Human bone marrow stem cells were cultured in 24 well culture dishes at an initial density of 25,000 cells/well. Cells were cultured for 0 14 days in DMEM supplemented with 10% FCS, 100U penicillin/mL, 100g streptomycin/mL, 0.2mM ascorbic acid, 10mM β-glycerophosphate, 2mM L-glutamine, 1mM sodium pyruvate and either 116, 164, 200, 225 or 240mg/L calcium chloride. Cultures at all calcium concentrations were grown without steroid hormone supplement, in the presence of 10nM Dexamethasone(Sigma). 1, 3, 7, 14 days after, alkaline phosphatase staining was done and total RNA was extracted, and the analysis of BMP-2, BMP-6, BSP and osteocalcin gene expression was done by RT-PCR.
Results and Conclusions: The results were as follows: 1.Calcium concentration affects the in vitro differentiation of human mesenchymal stem cell. 2. In the presence of dexamethasone, low calcium concentration favored osteoblastic gene expression.
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KEYWORD
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Bone marrow mesenchymal stem cell, Osteoblast, Osteogenic supplements, Calcium
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