体外团状培养系统中Ad-hTGF-β1诱导骨髓间充质干细胞向软骨细胞的分化  

Chondrocytic differentiation of bone marrow mesenchymal stem cells induced by Ad-hTGF-β1

作　　者：方军[1] 李华壮[1] 高克海[1] 陈景春[1] 

机构地区：[1]潍坊市益都中心医院骨科,山东省青州市262500

出　　处：《中国组织工程研究与临床康复》2009年第49期9717-9721,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:组织工程技术的发展给修复关节软骨缺损带来了希望,最近的比较研究证实骨髓间充质干细胞是修复全层软骨缺损的最佳细胞源。目的:在体外团状培养系统中,以Ad-hTGF-β1诱导骨髓间充质干细胞向软骨细胞定向分化,并对分化后的细胞进行鉴定。设计、时间及地点:以细胞为对象的重复观察测量实验,于2007-06/2008-01在解放军第三军医大学新桥医院中心实验室完成。材料:2月龄日本大耳白兔由解放军第三军医大学实验动物中心提供。方法:兔骨髓间充质干细胞体外分离培养扩增,Ad-hTGF-β1转染后,在团状培养系统中培养。通过组织学、免疫组化及反转录-聚合酶链反应方法检测其成软骨分化。主要观察指标:组织学染色观察细胞形态改变,免疫组化方法及反转录-聚合酶链反应检测蛋白多糖和Ⅱ型胶原的表达。结果:组织学染色显示,诱导后细胞呈软骨细胞样形态。免疫组化方法及反转录-聚合酶链反应结果显示,诱导后的细胞表达蛋白多糖和Ⅱ型胶原。结论:骨髓间充质干细胞经Ad-hTGF-β1诱导后在团状培养系统中可分化为软骨细胞。BACKGROUND：Tissue engineering development brings a hope for articular cartilage defect repair.Current studies have demonstrated that bone marrow mesenchymal stem cells （MSCs） are the best cell source to repair full-thickness cartilage defects.OBJECTIVE：To induce MSCs to differentiate into chondrocytes with Ad-hTGF-β1 in pellet culture system in vitro and identify the differentiated cells.DESIGN,TIME AND SETTING：Repetitive cellular measurements were performed at the Central Laboratory of Xinqiao Hospital,Third Military Medical University of Chinese PLA from June 2007 to January 2008.MATERIALS：Japanese rabbits,2 months old,were provided by the Laboratory Animal Center of Third Military Medical University of Chinese PLA.METHODS：The rabbit MSCs were isolated,cultured and expanded in vitro.After transfected with Ad-hTGF-β1,the cells were cultured in pellet culture system.Chondrogenic differentiation was evaluated by histological,immunohistochemical and RT-PCR techniques.MAIN OUTCOME MEASURES：Cell morphological changes were observed by histological staining;proteoglycan and type II collagen expression was detected by immunohistochemical and RT-PCR techniques.RESULTS：The induced cells exhibited a chondrocyte-like morphology by histological staining.Immunohistochemical and RT-PCR results showed that proteoglycan and type II collagen were expressed in the induced cells.CONCLUSION：Bone marrow MSCs cultured in pellet culture system can differentiate into chondrocytes under the induction of Ad-hTGF-β1.

关 键 词：骨髓间充质干细胞 转化生长因子Β 细胞分化 软骨细胞 

分 类 号：R394.2[医药卫生—医学遗传学]