【作者】 林云锋；

【导师】 田卫东；

【作者基本信息】 四川大学 ， 口腔临床医学， 2006， 博士

【摘要】 在临床医学中，因创伤、感染、肿瘤术后及先天缺陷导致组织缺损的修复重建，一直是困扰各专业医生的棘手难题。当前临床上使用的各种组织修复与重建方法虽然各有优缺点，但总体来说不尽人意。组织工程是应用细胞生物学和工程学的原理，对病损组织结构、功能的修复与重建进行研究开发的一门新兴科学。种子细胞是组织工程的首要因素，脂肪基质细胞(Adipose tissue-derived stromal cells，ASCs)是近年来研究最热门的种子细胞，被认为是最有可能用于临床的种子细胞之一。ASCs具有向脂肪细胞、成骨细胞、成软骨细胞和成肌细胞等多向分化的能力，加之脂肪贮备量大、取材方便的优势，使其更具实际的应用价值。 本课题采用酶消化法获取绿色荧光蛋白(Green fluorescent protein，GFP)转基因小鼠脂肪基质细胞，通过体外多向诱导的培养方法，研究内源性GFP对ASCs分化能力的影响；然后以腺病毒(Adenoviral vectors，Ad)为载体，转染外源性GFP基因到Balb／c小鼠ASCs中，比较内、外源性GFP对小鼠ASCs分化能力的影响；进而研究外源性GFP对人脂肪基质细胞分化能力的影响。在此基础上，结合组织工程的方法，研究ASCs体外成软骨分化能力，并使用经过成软骨诱导的ASCs复合藻酸盐支架材料，种植于裸鼠皮下，研究ASCs形成异位软骨组织的能力；再使用经过成骨诱导的ASCs复合纳米双相磷酸钙陶更多还原

【Abstract】 Current tissue-engineering approaches are being developed to repair and restore the function of damaged or diseased tissues. The basic principle involves an appropriate cell source and a biocompatible and biodegradable scaffold. Adipose tissue is derived from the embryonic mesenchyme and contains a stroma, in which the ASCs can be induced to multiple lineages in specific culture system similar to bone marrow stem cells (BMSCs). Compared with BMSCs, ASCs are easier to obtain, carry relatively lower donor site morbidity, and are available in large numbers of stem cells at harvest. There is less cell heterogeneity in ASCs, but apparent heterogeneity in BMSC. So it is suggested that population of ASCs represents a promising alternative for BMSCs used in cell-based musculoskeletal tissue-engineering strategies.In the part one, we aimed to confirm the trans-germ plasticity of endogenously GFP marked ASCs from GFP transgenic mice. The study was focused on two aspects. The first one was to determine whether GFP transgenic ASCs still could be induced to trans-germ multiple lineages in vitro. The second was aimed at the verification that the inductive conditions and the progress of terminal differentiation had no significant influence on GFP expression. We showed that ASCs isolated from transgenic mice have the capability to differentiate into adipogenic, osteogenic,更多还原