摘要目的 观察大鼠脂肪干细胞(adipose-derived stem cell,ASC)在聚乳酸-羟基乙酸共聚物(polylactic-co-glycolic acid,PLGA)支架上的生长状态、分裂增殖能力及黏附性,为进一步构建组织工程化鼓膜修复材料提供实验参考.方法 以Wistar大鼠为研究对象,分离其ASC,并与PLGA支架材料复合培养.分别行细胞波形蛋白(Vimentin)和Ki67免疫荧光染色,激光共聚焦显微镜下观察ASC在PLGA支架上的生长状态及分裂增殖能力.扫描电镜观察ASC与PLGA的黏附性.结果 Vimentin免疫荧光结果显示ASC在PLGA支架材料中生长状态良好,排列有序；ASC在PLGA支架上能正常活跃增殖,Ki67平均阳性指数((x)±s)为(8.21±1.76)％,对照组(未放支架材料,直接生长的ASC) Ki67平均阳性指数为(9.06±1.95)％,两组间差异无统计学意义(=1.03,P=0.30).扫描电镜结果显示,ASC与支架材料有较好的黏附性,伪足清晰可见,增殖的细胞间连接紧密.结论 大鼠ASC与PLGA有较好的相容性,可以在PLGA上正常黏附、增殖.

abstractsObjective To explore the morphology.,proliferative activity and adhesion of adiposederived stem cells (ASC) seeded in the polylactic-co-glycolic acid (PLGA) scaffold,and to provide experimental data for fabricating tissue engineered tympanic membrane repairment materials.Methods Wistar rats were selected,and the ASC were isolated and co-cultured with the PLGA scaffold. The morphology and proliferative activity of ASC in the scaffold were examined by immunofluorescence of Vimentin and Ki67 respectively.All the immunofluorescence signals were analyzed by a confocal laser scan microscopy system FLUOVIEW FV1000.The adhesion of ASC to the PLGA scalfold was determined by scanning electron microscopy.Results Immunofluorescence of vimentin showed rats ASC displayed normal morphology and grew orderly in the PLGA scalfold.Immunofluorescence of Ki67 showed the normal active proliferation of ASC in the scaffolds.The Ki67 mean positive index ( x ± s) of the ASC in the scalfold was (8.21 ± 1.76 ) ％,while in control group ( cultured without PLGA scalfold ) was ( 9.06 ± 1.95 ) ％.There was no statistic significance between the two groups ( t =1.03,P =0.30 ).Scanning electron microscopy demonstrated ASC adhered well to the PLGA scalfold,the pseudopodia of ASC could also be observed and the proliferative cell conjunction was tight.Conclusion ASC has good biocompatible to the PLGA scaffold and could normally adhere and proliferate in PLGA scaffold.