中文摘要：

目的 探索体外诱导人脂肪间充质干细胞成管状结构的可行性,为体外三维重建腺管样结构提供理论基础.方法 体外分离培养脂肪间充质干细胞,采用流式细胞仪检测细胞表面标记CD29、CD31、CD34、CD45、CD90、CD105的表达.经成骨诱导液（体积分数为10%胎牛血清,0.1μmol/L地塞米松,200μmol/L抗坏血酸,10mmol/L β-甘油磷酸钠,DMEM培养液）、成脂诱导液（体积分数为10%胎牛血清,1μmol/L地塞米松,200μmol/L吲哚美辛,0.5mmol/L IBMX,10mg/L胰岛素的DMEM培养液）、体外诱导脂肪间充质干细胞向骨细胞、脂肪细胞分化,并分别经碱性磷酸酶染色、油红O染色和鉴定.取培养至第3代的ADSCs,经含40ng/ml HGF 体积分数为10%胎牛血清 DMEM 诱导条件培养液,诱导ADSCs 在 Matrigel 胶中三维重建管状结构.结果 体外培养的人脂肪间充质干细胞呈梭形,生长增殖旺盛.流式细胞仪检测结果显示阳性表达：CD29、CD90、CD105阳性率分别为86.4%、 97.7%、89.8%;CD31、CD34、CD45阳性率分别为4.1%、3.7%、2.3%,经成脂、成骨定向诱导分化后,细胞分别呈现脂肪细胞、骨细胞的表型特征.经40ng/mlHGF诱导培养5d后可在Matrigel胶中呈现管状结构特征.结论 脂肪间充质干细胞可望用于组织工程研究构建腺管样结构.

英文摘要：

Objective To explore the feasibility of tubular structure inducted by human adipose derived mesenchymal stem cells （ADSCs）, which provide the theoretical basis for the three-dimensional reconstruction of duct-like structures in vitro. Methods ADSCs were isolated and cultured in vitro, and expression of CD29, CD31 and CD34, CD45, CD90, CD105 in ADSCs were detected with flow cytometry. ADSCs were induced into osteocytes and lipoeytes by osteogenic induetion medium （ 10% fetal bovine serum, 0. 1 μmol/L dexametha- sone, 200/.Lmol/L ascorbie acid, 10 mmoL/L β-glyceropbospbate, DMEM culture medium）and adipogenic induction medium （10% fetal bovine serum, 1 μmol/L, dexametbasone, 200 μmol/L indomethaein, 0.5 mmol/L IBMX, 10 mg/L insulin in DMEM culture medium） in vitro, respectively and then identificated by alkaline pbosphatase staining and oil red O staining. The third generation of cultured ADSCs treated with DMEM conditioned medium （10% fetal bovine serum and 40 ng/ml HGF ）were inducted into the three-dimensiona tubular structures in Matrigel glue. Results The cultured human ADSCs proliferated well with fusiform shape. Flow cytometry results showed rates of positive expression of CD29, CD90 and CD105 were 86.4%, 97.7%, 89.8%, respectively; the positive rates of CD31, CD34, CD45 expression were 4. 1%, 3.7%, 2.3% ,respectively. The induced cells were presented the phenotypic characteristics of the lipoeytes and bone osteoeytes which showed tubular structures at 5 days after treating by HGF in Matrigel. Conclusion ADSCs could be used in tissue engineering research, like constructing glandular duct-like structures.