摘要
背景:促甲状腺素、胰岛素或类胰岛素生长因子等对甲状腺细胞的发育分化、特异基因的表达及功能的产生等起着重要作用,故实验模拟甲状腺的体内胚胎发生过程逐步加入以上刺激因子对骨髓间充质干细胞进一步诱导。目的:探讨在特定环境下体外定向诱导成人骨髓间充质干细胞分化为甲状腺细胞的可行性。方法:采用密度梯度法分离培养骨髓间充质干细胞,应用促甲状腺素、胰岛素等试剂进行诱导。利用倒置光学显微镜、透射电镜观察细胞分化过程的形态学变化,免疫荧光等检测方法研究成人骨髓间充质干细胞的分化情况。结果与结论:诱导组培养第3天可见骨髓间充质干细胞由长梭形变为圆形、椭圆形或三角形,不规则形生长,边界欠清晰。从第8天开始可见贴壁的胚胎体逐渐呈扁平状。诱导培养第7天可见分化细胞中有甲状腺细胞的特有基因如TSHr的表达;第9天检测到分化细胞中甲状腺细胞标记物TTF-1的表达。结果初步表明骨髓间充质干细胞可以在体外诱导分化为甲状腺细胞,是研究甲状腺疾病组织工程治疗的理想种子细胞。
BACKGROUND：Thyrotrophin,insulin or insulin-like growth factor play important roles in development,differentiation,heterogenic expression and function production of thyroid cells.Thus,this study simulated thyroid in vivo embryogenesis,and gradually added above-mentioned stimulating factor to further induce bone marrow mesenchymal stem cells（BMSCs）.OBJECTIVE：To investigate the feasibility of differentiation of the adult BMSCs into thyroid cells under specific circumstances in vitro.METHODS：BMSCs were isolated by the density gradient method.BMSCs were induced with thyrotrophin and insulin.Morphological changes were observed by the inverted optical microscope and transmission electron microscopy.Immune fluorescence detection method was used to differentiate adult BMSCs.RESULTS AND CONCLUSION：On day 3 following culture in the induction group,BMSC morphology changed from spindle to round,ellipse or triangle,grew irregularly,with clear boundary.On day 8,adherent embryo became flat.On day 7,differentiated cells expressed thyroid cell gene TSHr.On day 9,differentiated cell markers TTF-1 expression was detected in thyroid cells.Results suggested that BMSCs can be induced to differentiate into thyroid cells in vitro and are ideal seed cells in the study of tissue engineering treatment of thyroid diseases.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2011年第6期951-954,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
