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.