大鼠牙髓干细胞和骨髓间充质干细胞分化成骨样细胞能力对比研究.doc

大鼠牙髓干细胞和骨髓间充质干细胞分化成骨样细胞能力对比研究 　　[摘要]目的：以骨髓间充质干细胞作为参考，讨论牙髓干细胞作为骨组织修复种子细胞的可行性。方法：通过酶消化法获得大鼠牙髓干细胞，离心法获得骨髓间充质干细胞，贴壁培养，通过倒置光学显微镜观察二者形态差异；流式细胞技术鉴定骨髓间充质干细胞的间质细胞表面标志物表达；ＭＭＴ法检测细胞生长曲线；特定的成骨诱导液诱导干细胞成骨分化，免疫荧光法检测成骨细胞表面标志物表达。结果：分离的大鼠骨髓间充质干细胞与牙髓干细胞形态学以及生长特性具有相似性，且符合间充质干细胞特性，牙髓干细胞第4天进入对数生长期，第7天进入平台期，骨髓间充质干细胞第4天进入对数生长期，第8天进入平台期；经过成骨诱导后二者都具备成骨样细胞分化能力，牙髓干细胞在成骨诱导后的骨钙素表达上与骨髓间充质细胞有着相似的能力；牙本质泌涎蛋白表达阳性。结论：牙髓干细胞具有与骨髓间充质干细胞都具有成骨分化能力，但牙髓干细胞细胞成分相对复杂，增殖能力较强。 　　[关键词]牙髓干细胞；培养；诱导；成骨样细胞 　　[中图分类号]Q813.1[文献标识码]A[文章编号]1008-6455（2011）04-0597-03 　　 　　Comparative study on the capacity of rat dental pulp stem cells and bone marrow mesenchymal stem cells in differentiating to osteoblast-like cells 　　ZHAI Xu,CAO Rui,CAI Lei,WU Jing-guo,SUN Xue-jian,LV Chang-sheng 　　(Research Center of Plastic Surgery Hospital,Chinese Academy of Medical Sciences，Peking Union Medical College,Beijing 100144,China) 　　 　　AbstractObjectiveTo research the capacity of dental pulp stem cells which was induced to osteoblast-like cells , by comparing bone marrow mesenchymal stem cells. Methods Pulps and bone marrow were isolated from 6-weeks rats,then pulps was been digested by collagenase I. morphology difference was distinguished by optical microscope. FCM(flow cytometry) was uesd for identifying the purity of BMSCs. Drawing growth curves for cells was chooen. Cells was introduced by osteogenetic induced flow. OCN and DSP were targeted in immunofluorescence test, as being oseteo-markers.ResultsCells isolated from rats tissues were corresponded to the mesenchymal cell. Cells proliferated faster at the fourth day after cultivation.Both of the tow cells have the capacity of differentiating to osteoblast-like cells though inducing cultivation. OCN and DSP were detected on both cells.ConclusionDPSCs had the same capacity to differentiate to osteoblast-like cells with BMSCs,and got a more ability of proliferation. 　　Key words:dental pulp stem cells;cultivation;inducing;osteoblast-like cells 　　 　　骨组织缺损修复研究是近年来研究较多的热门学科，越来越多的研究者