甲状腺素诱导人脐血间充质干细胞向软骨细胞分化中的作用  

Thyroxine effects on induced differentiation of human umbilical cord blood-derived mesenchymal stem cells into chondrocytes

作　　者：杨涛[1] 江波[1] 徐鹏[1] 林刚[1] 

机构地区：[1]南京医科大学附属南京儿童医院,江苏省南京市210008

出　　处：《中国组织工程研究》2015年第1期7-11,共5页Chinese Journal of Tissue Engineering Research

基　　金：南京市卫生局医学科技发展课题(YKK12112)~~

摘　　要：背景:研究发现共培养可诱导人源干细胞向特定细胞分化,但如何获得更多的组织工程所需的种子细胞是研究中的难题。目的:探讨不同浓度甲状腺素在人脐血间充质干细胞向兔软骨细胞分化中的作用。方法:将人脐血间充质干细胞与兔软骨细胞按照2∶1比例共培养,并用含有不同浓度甲状腺素(0,0.01,0.1,1,10μmol/L)的培养基分组刺激,共培养14 d后分别观察各组细胞形态,并提取细胞RNA和蛋白质,Real-time PCR检测聚集蛋白多糖m RNA及Ⅱ型胶原m RNA表达量,Western blotting技术检测Ⅱ型胶原及聚集蛋白多糖蛋白表达水平。结果与结论:加入0.01,0.1,1,10μmol/L的甲状腺素干预后,聚集蛋白多糖、Ⅱ型胶原m RNA和蛋白的表达水平均随甲状腺素浓度增加而增强,与单纯共培养组比较差异有显著性意义(P<0.05)。实验证明高浓度甲状腺素可增强人脐血间充质干细胞与兔软骨细胞共培养后向软骨细胞分化的能力。BACKGROUND：Human umbilical cord blood-derived mesenchymal stem cels can be induced through the co-culture to differentiate into other cels, but how to get more seed cels for tissue engineering is one of the most difficult problems. OBJECTIVE： To investigate the role of the different concentrations of thyroxine in chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cels by co-culture with rabbit chondrocytes. METHODS：Human umbilical cord blood-derived mesenchymal stem cels were co-cultured with rabbit chondrocytes at 2：1, and stimulated by medium containing different concentrations of thyroxine （0, 0.01, 0.1 and 1, 10 μmol/L）. Co-cultured cels with no thyroxine served as control group. After 14 days of co-culture, the cel RNA and protein were extracted, mRNA expressions of aggrecan and colagen type II were detected by real-time PCR, and protein expression of aggrecan and colagen type II were detected by western blot assay. RESULTS AND CONCLUSION：After intervention with 0.01, 0.1, 1, 10 μmol/L thyroxine, the mRNA and protein expressions of aggrecan and colagen type II were enhanced with the increase of thyroxine concentration, which were significantly different from those in the control group （P〈 0.05）. Experimental findings indicate that high levels of thyroxine can enhance the chondrogenic ability of human umbilical cord blood-derived mesenchymal stem cels co-cultured with rabbit chondrocytes.

关 键 词：胎血 间质干细胞 软骨细胞 共同培养技术 甲状腺素 干细胞 脐带脐血干细胞 共培养 细胞分化 Ⅱ型胶原 聚集蛋白多糖 软骨组织工程 种子细胞 

分 类 号：R394.2[医药卫生—医学遗传学]