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目的 通过对骨髓间充质干细胞 (mesenchymalstemcell,MSC)向成骨细胞系转化过程中表征变化进行观察 ,以期更全面地了解MSC在体外的成骨分化过程。方法 将羊MSC进行体外诱导分化 ,进行组织学、免疫组化、X线衍射分析等检测及原子力显微镜观察。结果 诱导后的MSC碱性磷酸酶染色阳性率达 90 %以上 ,Ⅰ型胶原免疫组化和钙结节染色均呈阳性 ;诱导后细胞形成的结节样物中主要由羟基磷灰石组成 ;原子力显微镜观察见诱导后的MSC具有成熟的成骨细胞特性。结论 体外培养的MSC能被诱导成骨 ;运用原子力显微镜可以清晰地看到MSC在向成骨细胞分化过程中表征的变化。
OBJECTIVE: To investigate the characterization of mesenchymal stem cells (MSC) into osteoblast cell line in order to obtain a more complete understanding of the osteogenic differentiation process of MSC in vitro. Methods Sheep MSC were induced and differentiated in vitro and examined by histology, immunohistochemistry, X-ray diffraction and atomic force microscopy. Results The positive rate of alkaline phosphatase staining was more than 90% after induction. The immunohistochemical staining of type Ⅰ collagen and calcium nodules showed positive staining. The nodules formed after induction were mainly composed of hydroxyapatite. Atomic force microscopy showed that the induced MSCs have mature osteoblastic properties. Conclusion MSCs cultured in vitro can be induced into osteoblasts. The changes in the characterization of MSC during differentiation into osteoblasts can be clearly seen by atomic force microscopy.