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目的 :用人骨形态发生蛋白 2腺病毒表达载体 (Ad- BMP- 2 )转染兔骨髓基质干细胞 (BMSC) ,观察Ad- BMP- 2转染对细胞分化的影响。方法 :将 Ad- BMP- 2转染体外培养的兔 BMSC,7d后利用原位杂交和免疫组化方法检测细胞内 BMP- 2及 型胶原的表达 ,同时行碱性磷酸酶染色。结果 :BMP- 2和 型胶原原位杂交法显示实验组细胞胞浆中有棕黄色阳性杂交信号 ,对照组阴性。 BMP- 2和 型胶原免疫组化法显示实验组细胞胞浆棕黄色阳性着色 ,胞核阴性表达 ,对照组阴性 ;碱性磷酸酶染色见实验组细胞胞浆呈棕黑色阳性着色 ,可见细小的棕黑色颗粒 ,对照组染色阴性。结论 :Ad- BMP- 2可高效转染 BMSC,且促进其向成骨细胞和软骨细胞方向转化。
OBJECTIVE: To transfect rabbit bone marrow stromal stem cells (BMSCs) with human BMP-2 adenovirus vector (Ad-BMP-2) and observe the effect of Ad-BMP-2 transfection on cell differentiation. METHODS: Ad-BMP-2 was transfected into rabbit BMSCs cultured in vitro. After 7 days, the expression of BMP-2 and collagen in cells was detected by in situ hybridization and immunohistochemistry. Meanwhile, alkaline phosphatase staining was performed. RESULTS: BMP-2 and collagen in situ hybridization showed brown-yellow positive hybridization signal in the cytoplasm of the experimental group, but negative in the control group. Immunohistochemistry of BMP-2 and type collagen showed that the cytoplasm of the experimental group showed brownish-yellow staining and negative staining of the cytoplasm, negative in the control group; alkaline phosphatase staining showed that the cytoplasm of the experimental group showed a brownish-black staining, Brown black particles, the control group stained negative. CONCLUSION: Ad-BMP-2 can efficiently transfect BMSC and promote its transformation to osteoblasts and chondrocytes.