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腺病毒介导的hTGF-β2转移诱导预分化的人脂肪来源干细胞形成异位新软骨。

Ectopic neocartilage formation from predifferentiated human adipose derived stem cells induced by adenoviral-mediated transfer of hTGF beta2.

作者信息

Jin Xiao bing, Sun Yong sheng, Zhang Ke, Wang Jing, Shi Tai ping, Ju Xiao dong, Lou Si quan

机构信息

Orthopaedics Department, Peking University, Third Hospital, Beijing 100083, PR China.

出版信息

Biomaterials. 2007 Jul;28(19):2994-3003. doi: 10.1016/j.biomaterials.2007.03.002. Epub 2007 Mar 14.

Abstract

Chondrogenic potential of human adipose derived stem cells (hASCs) makes them a possible source of seeding cells for cartilage tissue engineering. In this study, chondrogenic differentiation of hASCs induced by transduction with replication-deficient adenovirus carrying human transforming growth factor beta2 (Ad5-hTGF beta2) was demonstrated by RT-PCR, immunohistochemistry staining, biochemical and western blot analysis. To evaluate if the in vitro differentiated hASCs could keep their chondrocytic phenotype and produce neo-cartilage in vivo, predifferentiated hASCs were seeded in different scaffolds and implanted in subcutaneous pockets on the dorsum of nude mice. After 4 and 12 weeks culture in vivo, specimens were harvested and examined by histological and immunohistochemical analysis, cartilage-like tissue formation was only found in alginate gel and PLGA/alginate compound groups, in PLGA group, fibrous tissues and angiogenesis ingrowth were observed. These findings demonstrated that adenovirus-mediated hTGF beta2 gene transfer could induce hASCs into a chondrogenic lineage in vitro, however, this predifferentiation did not guarantee ectopic cartilage formation in vivo unless appropriate three-dimensional scaffolds were used as the cell carry vehicles.

摘要

人脂肪来源干细胞(hASCs)的软骨生成潜能使其成为软骨组织工程种子细胞的一个可能来源。在本研究中,通过逆转录聚合酶链反应(RT-PCR)、免疫组织化学染色、生化分析和蛋白质印迹分析,证实了携带人转化生长因子β2(Ad5-hTGFβ2)的复制缺陷型腺病毒转导诱导hASCs的软骨分化。为了评估体外分化的hASCs是否能在体内保持其软骨细胞表型并产生新软骨,将预分化的hASCs接种到不同支架中,并植入裸鼠背部的皮下囊袋。在体内培养4周和12周后,收集标本并进行组织学和免疫组织化学分析,仅在藻酸盐凝胶和聚乳酸-羟基乙酸共聚物/藻酸盐复合组中发现了软骨样组织形成,在聚乳酸-羟基乙酸共聚物组中,观察到纤维组织和血管内生。这些发现表明,腺病毒介导的hTGFβ2基因转移可在体外诱导hASCs向软骨谱系分化,然而,除非使用合适的三维支架作为细胞载体,这种预分化并不能保证在体内形成异位软骨。

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