中文摘要：

目的探讨并建立一种简便有效的体外分离纯化及扩增大鼠骨髓间质干细胞的方法，并研究其生物学特性，对其进行初步鉴定，为进一步诱导分化为胃黏膜上皮细胞奠定基础。方法利用贴壁培养法分离纯化SD大鼠骨髓间质干细胞，体外培养，传代扩增，在倒置显微镜下连续观察细胞的形态变化，并用免疫组化方法鉴定骨髓间质干细胞表面CD29，CD44，CD34的表达。将加入成骨、成脂肪诱导剂的骨髓间质干细胞体外培养2～3周，观察细胞形态变化，并做油红0染色，鉴定成骨及成脂肪分化的结果。结果原代培养8—10d后可分离得到骨髓间质干细胞，且骨髓间质干细胞体外培养生长状况良好，呈均一的成纤维细胞样，传代周期为3～4d。免疫组化显示90％以上的骨髓间质干细胞CD29、CD44阳性，CD34阴性，且可以将其定向诱导为脂肪细胞和成骨细胞。结论利用贴壁培养法可分离培养出大量大鼠骨髓间质干细胞，且此方法简便易行。

英文摘要：

Objective To establish a simple and effective method of isolation, purification and culture of rat bone marrow mesenchymal stem cells （MSC） in vitro, and to explore its biological characteristics and identify the cells in order to provide evidence for gastric epithelial cells differentiation. Methods MSCs of Sprague Dawley （SD） rat were isolated, purified, subcultured and amplified by means of adherence culture. Cell growth was observed under an inverted microscope and membrane antigen CD29, CD44, CD34 were detected through immunohistochemistry. Osteoblastic differentiation and adipocytic differentiation were induced by different inducers for 2 - 3 weeks in vitro. Morphology changes were observed and MSCs were stained with oil red O after 3 weeks to verify the results of differentiation. Results MSCs could be separated 8 - l0 days after primary culture and morphology of MSCs was identical fibroblast-like. MSCs were passaged every 3 - 4 days. Immunohistochemistry staining showed that CD29, CD44 were positive and CD34 was negative in more than 90% MSCs. They could be induced to differentiate into adipoctyes and osteoblasts. Conclusion Adherence culture is a simple and practical method for separated and amplified MSCs.