CAJ | 학술논문

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.
배경:조직공정기술적발전급수복관절연골결손대래료희망,최근적비교연구증실골수간충질간세포시수복전층연골결손적최가세포원.목적:재체외단상배양계통중,이Ad-hTGF-β1유도골수간충질간세포향연골세포정향분화,병대분화후적세포진행감정.설계、시간급지점:이세포위대상적중복관찰측량실험,우2007-06/2008-01재해방군제삼군의대학신교의원중심실험실완성.재료:2월령일본대이백토유해방군제삼군의대학실험동물중심제공.방법:토골수간충질간세포체외분리배양확증,Ad-hTGF-β1전염후,재단상배양계통중배양.통과조직학、면역조화급반전록-취합매련반응방법검측기성연골분화.주요관찰지표:조직학염색관찰세포형태개변,면역조화방법급반전록-취합매련반응검측단백다당화Ⅱ형효원적표체.결과:조직학염색현시,유도후세포정연골세포양형태.면역조화방법급반전록-취합매련반응결과현시,유도후적세포표체단백다당화Ⅱ형효원.결론:골수간충질간세포경Ad-hTGF-β1유도후재단상배양계통중가분화위연골세포.
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-pi 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 transacted 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.