摘要
背景:有研究发现关节软骨微环境可诱导骨髓间充质干细胞向软骨细胞分化。目的:验证软骨细胞能否诱导脂肪干细胞向软骨细胞分化,以便成为组织工程构建关节软骨可能的新方法。设计、时间及地点:随机对照细胞实验,于2007-09/2008-07在南京大学生物医药生物技术国家重点实验室完成。材料:清洁级成年新西兰大白兔,雌雄不限,体质量1.5～2.0kg。方法:分离培养原代新西兰大白兔脂肪干细胞和软骨细胞,分别种植于6孔板和插入式细胞培养皿共培养2周后,胰酶消化终止。脂肪干细胞分别行油红O染色、碱性磷酸酶检测和甲苯胺蓝染色法鉴定。主要观察指标:倒置显微镜观察共培养前后脂肪干细胞的形态变化。甲苯胺蓝染色法检测共培养后脂肪干细胞的蛋白多糖的表达水平。免疫细胞化学检测共培养后脂肪干细胞的Ⅱ型胶原的表达水平。反转录-聚合酶链反应检测蛋白多糖和Ⅱ型胶原的基因转录水平。结果:脂肪干细胞分别经油红O染色、碱性磷酸酶染色和甲苯胺蓝染色法鉴定证实,分离的细胞具有多向分化能力,是脂肪来源的间充质干细胞。共培养1周后部分脂肪干细胞变圆,2周时其蛋白多糖和Ⅱ型胶原的基因转录和蛋白表达均增高。结论:与软骨细胞共培养后,脂肪干细胞可以被诱导成软骨样细胞。
BACKGROUND： Study shows that bone marrow mesenchymal stem cells can be induced into chondrocytes in the microenvironment of articular cartilage. OBJECTIVE： To verify the feasibility of inducing adipose derived stem cells （ADSCs） into chondrocytes so as to provide a novel approach for articular cartilage tissue engineering. DESIGN, TIME AND SETTING： A randomized cell trial was performed at the State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University from September 2007 to July 2008. MATERIALS： Adult New Zealand white rabbits of clean grade, weighing 1.5 to 2.0 kg, of either gender. METHODS： Primary ADSCs and aricular chondrocytes from New Zealand white rabbits were isolated and then seeded on 6-pore plates and millicell culture plate inserts separately. After 2 weeks of co-culture, trypsinization was terminated and ADSCs were detected with oil red O staining method, alkaline phosphatase detection method and toluidine blue staining method respectively. MAIN OUTCOME MEASURES： The cytomorphology of ADSCs was observed under inverted microscope before and after co-culture respectively. After co-culture, the proteoglycan expression of ADSCs was detected with toluidine blue staining method and the collagen type Ⅱ expression with immunocytochemistry detection method. The Genetic transcription level of aggrecan and collagens type Ⅱ was determined with reverse transcriptase-polymerase chain reaction. RESULTS： All of the three detection method （namely, oil red O staining method, alkaline phosphatase detection method and toluidine blue staining method） proved that ADSCs had the multi-directional differentiation ability and were mesenchymal stem cells derived from adiposes. After 1 week of co-culture, parts of ADSCs were rounded morphologically. The genetic transcription level and the expression of both aggrecan and collagens type Ⅱ increased after 2 weeks of co-culture. CONCLUSION： ADSCs are able to be induced into cartilage-like cells when being co-cultured with chondrocytes.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第24期4632-4636,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
作者简介
张勇，男，1973年生，安徽省当涂县人，汉族，解放军第二军医大学骨科在读博士，主治医师，主要从事关节外科与组织工程研究。
通讯作者：赵建宁，硕士，教授，博士生导师，解放军第二军医大学南京临床医学院，解放军南京军区南京总医院骨科，江苏省南京市210002zhaojianning．0207⑥163．com
