中文摘要：

研究表明脂肪组织是多潜能干细胞的又一新来源,脂肪组织来源的干细胞可被用于细胞治疗及组织工程。然而,传统的培养方法很难满足临床需求。为获得大量的脂肪干细胞以满足临床需求,现将脂肪干细胞接种到胶原/壳聚糖支架上,比较细胞在静态环境和在转瓶中动态扩增的情况。通过CCK-8检测细胞增殖情况;并分析葡萄糖和乳酸的代谢情况;14d后扫描电镜观察细胞在支架内的生长情况;流式细胞仪检测干细胞相关表面标记表达;RT-PCR检测干细胞相关转录因子的表达;并检测扩增后的干细胞的多向分化潜能。结果表明：在动态微环境中扩增14d后,与静态条件下相比,支架内的细胞具有更强的增殖活性和更好的多向分化潜能。所扩增的细胞能够保持原有干细胞的特性。结论：所设计的支架-转瓶培养系统是一个简便有效的扩增脂肪干细胞的方法。

英文摘要：

Recent reports indicate that adipose tissue is a novel source of multipotent stem cells, which can be used in cell therapy and tissue engineering. However, traditional cultivation of adipose tissue-derived stem cells （ADSCs） is hard to meet the need of clinical applications. In order to obtain a great number of ADSCs, we seeded ADSCs in collagen/chitosan scaffolds and compared the proliferation of ADSCs in three dimensional （3D） static environment in dishes and 3D dynamic environment in spinner flask. The growth dynamic parameters of ADSCs were examined by CCK-8 kit every other day; variations of glucose and lactic acid concentration were analyzed every day. After 14 days, the cells were observed with scanning electron microscope （SEM）. The surface markers, the specific transcription factors and the multi-differentiation potential were also assayed to identify the stemness of expanded cells. The results show that in spinner flask, the cells in scaffolds could be expanded by more than 26 times in 14 days, and they present better morphology and vitality and stronger differentiation ability than the cells cultivated in scaffolds statically. All cells maintain stem cell characteristics after proliferation. Therefore, spinner flask cultivation is an easy-to-use inexpensive system for expanding ADSCs in 3D scaffolds.