胰岛素样生长因子1对人脂肪来源的间充质干细胞向软骨细胞定向诱导分化的作用  

Insulin-like growth factor-1 effects on directional differentiation of human adipose-derived mesenchymal stem cells into chondrocytes

作　　者：周全[1] 邓展生[2] 朱勇[2] 李宝军[2] 张少先[1] 赵加力[1] 

机构地区：[1]徐州医学院附属淮安医院骨科,江苏省淮安市223002 [2]中南大学湘雅医院脊柱外科,湖南省长沙市410008

出　　处：《中国组织工程研究与临床康复》2010年第10期1785-1790,共6页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:近年研究者发现胰岛素样生长因子1还可以诱导骨髓来源间充质干细胞向软骨细胞方向分化,但目前尚未见胰岛素样生长因子1诱导脂肪来源间充质干细胞向软骨细胞方向分化及在此过程中与转化生长因子β1相互作用的报道。目的:观察胰岛素样生长因子1诱导脂肪来源间充质干细胞向软骨细胞定向分化的可能性及在诱导分化中和转化生长因子β1的相互作用。方法:获取脂肪来源间充质干细胞,以2×105cells/cm2的密度接种于培养瓶,使用含胰岛素样生长因子1或(和)转化生长因子β1的无胰岛素软骨诱导剂诱导脂肪来源间充质干细胞。2周后获取细胞,制备细胞爬片,进行甲苯胺蓝染色和Ⅱ型胶原免疫组织化学染色,观察细胞内高硫酸化的蛋白聚糖和Ⅱ型胶原着色情况。RT-PCR检测Ⅱ型胶原蛋白、aggrecan及Sox9mRNA的表达。结果与结论:加入诱导剂后,甲苯胺蓝染色显示3个诱导组细胞呈多角形,胞浆及胞膜呈蓝色异染。Ⅱ型胶原免疫组织化学染色显示3个诱导组细胞胞浆及胞膜呈棕黄色着色。RT-PCR检测显示胰岛素样生长因子+转化生长因子组Ⅱ型胶原蛋白、aggrecan、Sox9mRNA的表达均显著强于胰岛素样生长因子组和转化生长因子组,胰岛素样生长因子组和转化生长因子组显著强于对照组,而胰岛素样生长因子组与转化生长因子组差异无显著性意义。提示胰岛素样生长因子1可以单独诱导脂肪来源间充质干细胞向软骨细胞分化,表达软骨细胞特异性细胞表型,胰岛素样生长因子1与转化生长因子β1诱导脂肪来源间充质干细胞向软骨细胞分化有协同作用。BACKGROUND： Recently, researches have found that insulin-like growth factor-1 （IGF-1） can induce the differentiation of bone marrow-derived mesenchymal stem cells （BMSCs） into chondrocytes, but there are no reports concerning the differentiation of adipose-derived mesenchymal stem cells （ADMSCs） into chondrocytes induced by IGF-1, as well as interaction with transforming growth factor-β1 （TGF-β1） during this process. OBJECTIVE： To explore the possibility of inducing ADMSCs chondrogenic differentiation by using IGF-1 and the interaction with TGF-β1 in induction. METHODS： ADMSCs were obtained, and seeded at 2×105cells/cm2 in culture flask. Insulin-free chondrogenic media containing IGF-1 or （and） TGF-β1 were used to induce ADMSCs. 2 weeks later, cells were harvested and stained by using toluidine blue and collagen Ⅱ antibody immunohistochemistry. Intracellular sulfated proteoglycan and collagen Ⅱ coloring were observed. Reverse transcription-polymerase chain reaction （RT-PCR） was used to detect the expression of collagen Ⅱ, aggrecan and Sox9 mRNA. RESULTS AND CONCLUSION： After induced, toluidine blue stain exhibited that the cells in the three induction groups were polygonal, with cytoplasm and cell membrane of blue different dyeing. Immunohistochemistry for type II collagen demonstrated that cytoplasm and cell membrane were stained brown in three induction groups. RT-PCR revealed that the expression of collagen Ⅱ, aggrecan, Sox9 mRNA of IGF ＋ TGF group were significantly greater than the IGF and TGF groups, and IGF and TGF groups were significantly stronger than the control group. No significant difference was determined between the IGF and TGF groups. These results indicated that IGF-1 can induce chondrogenic differentiation from ADMSCs, expressing chondrocyte specific cell phenotype. There is synergism of IGF-1 and TGF-β1 to induce the differentiation of ADMSCs into chondrocytes.

关 键 词：脂肪间充质干细胞 细胞因子 组织工程 分化 软骨 

分 类 号：R394.2[医药卫生—医学遗传学]