论文部分内容阅读
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。
OBJECTIVE: To study the changes of mRNA expression of osteoblast differentiation-related genes ALP, OCN and COL I induced by goat bone marrow mesenchymal stem cells into osteoblasts. Methods: Bone marrow stromal stem cells were cultured by whole bone marrow culture method. Osteoblasts were induced by using osteogenic culture medium. Osteoblasts were identified by morphological observation, immunohistochemical staining and calcium nodules. RT- PCR and real-time quantitative PCR were used to detect the expression of ALP, OCN and COL I in bone marrow stromal stem cells after 2 weeks of induction. Uninduced bone marrow stromal stem cells served as the control group. RESULTS: Bone marrow stromal cells were successfully differentiated into osteoblasts by osteoblast identification. OCN and COL I genes were up-regulated in cell-induced group, and ALP gene expression was not significantly different between induced group and non-induced group. CONCLUSION: The goat BMSCs were successfully induced to differentiate into osteoblasts, and the related genes of osteogenic differentiation changed to adapt to the function of osteoblasts after induction.