山羊骨髓基质干细胞向成骨细胞诱导后成骨分化相关基因表达的变化  

The change of bone differentiation related gene expression after goat bone marrow mesenchymal stem cells inducing to osteoblast

作　　者：何悦[1] 贺捷[1] 邱蔚六[1] 张秀丽[1] 竺涵光[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院·口腔医学院口腔颌面外科,上海市口腔医学重点实验室,上海200011

出　　处：《中国口腔颌面外科杂志》2009年第6期545-550,共6页China Journal of Oral and Maxillofacial Surgery

基　　金：国家自然科学基金(30600714);上海市科学技术委员会启明星计划(07QA14039);上海市重点学科建设项目(S30206)~~

摘　　要：目的:研究山羊骨髓基质干细胞向成骨细胞诱导后成骨分化相关基因ALP、OCN、COL I mRNA表达水平的变化。方法:采用全骨髓培养法培养骨髓基质干细胞,使用成骨条件培养液体外诱导成骨细胞,通过细胞形态学观察、免疫组化染色、钙结节等检测手段进行成骨细胞鉴定,采用RT-PCR和实时定量PCR检测诱导2周后骨髓基质干细胞ALP、OCN、COL I的表达水平,未诱导的骨髓基质干细胞作为对照组。结果:通过成骨细胞鉴定,骨髓基质干细胞成功诱导分化为成骨细胞;OCN和COL I基因在细胞诱导组表达上调,ALP基因表达在诱导组和未诱导组间无显著差异。结论:成功诱导山羊BMSCs向成骨细胞分化,成骨分化相关基因在诱导后为适应成骨细胞的功能发生变化。PURPOSE： To study the bone differentiation related gene expression ALP, OCN, COL I changes in mRNA level after goat bone marrow rnesenchymal stem cells （BMSCs） inducing to osteoblast. METHODS： Whole bone marrow euhure method was used to amplify the BMSCs in vitro. Bone formation conditioned medium was adopted to induce the BMSCs differentiate to osteoblast. Characterization was performed by using cell morphology observation, immuno-histochemical staining and Von Kossa methods. RT-PCR and real-time PCR were used to examine the bone differentiation related gene OCN. eollagen I, ALP mRNA expression. The untreated BMSCs were used as the control. RESULTS： After osteoblast eharaelerization, the BMSCs were successfully differentiated to osteoblast. The OCN, collagen i mRNA expression were up-graded,while ALP mRNA expression had little change. CONCLUSION： BMSCs can be sueeessfully differentiated to osteoblast by using bone formation conditioned medium. The bone differentiation related genes change to aceonunodate the osteoblast funetion.

关 键 词：骨髓基质干细胞 实时定量PCR 成骨分化相关基因 羊 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]