West China Journal of Stomatology

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Cultivation and Induced Differentiation of Bone Marrow Stromal Cells of SD Rats with TypeⅠOsteoporosisin vitro

TANG You-chao, TANG Wei, TIANWei-dong, CHENXi-zhe, LISheng-wei   

  1. KeyLab. ofOralBiomedicalEngineering ofMin- istry ofEducation, Sichuan University, Chengdu610041, China
  • Received:2005-04-25 Revised:2005-04-25 Online:2005-04-20 Published:2005-04-20

Abstract:

Objective To study the biological features and osteoblast/adipocyte phenotypes of bone marrow stromal cells (BMSCs) of Sprague-Dawley (SD) rats withTypeⅠosteoporosis by induced culture.Methods Six-month-old SD rats were used in this study.16 female rats were randomly divided into two groups. eight rats were ovariectomied as experimental group to estab- lish the modle ofTypeⅠosteoporosis, while other rats received sham-operation. Three month later BMSCs of 16 rats were isolated by discontinueous gradient centrifugation and then plated inα-MEM medium as primary culture. Secondary harvested cells were cultured for 14 days inα-MEMmedium supplemented with dexamethasone, ascorbic acid, vitamin D3,β-glycerophosphate or dex- amethasone, 3-isobutyl-1-methylxanthine, insuline, and indomethine. The cells were screened by inverted microscope each day and cell growth was studied with cell counting. The osteoblast and adipocyte phenotypes were verified by cytochemistry staining, counted the percentage of positive stained cells.Results The weight and bone mineral density of rats were statistically different between experimental group and control group. Gomori and Von Kossa′s staining demonstrated positive osteoblast phenotypes of al- kaline phosphatase and mineralized nods by osteogenic inducer, while Oil Red O staining identified BMSCs treatedwith adipogenic medium resulted in adipocyte formation and there was no significant difference in the percentage of positive stained cells between two groups.Conclusion The model of TypeⅠosteoporosis has been established successfully. BMSCs from SD rats with osteo- porosis maintain their differentiation potential.

Key words: osteoporosis, marrow stromal cell, adult stem cell