外源性TGF-β1诱导大鼠骨髓间充质干细胞向肌成纤维细胞分化的实验研究

发布时间：2018-04-23 23:35

本文选题：颌骨 + 骨髓间充质干细胞　；参考：《口腔医学》2014年01期

【摘要】：目的探讨大鼠骨髓间充质干细胞BMSCs(bone marrow mesenchymal stem cells)的分离、培养、扩增方法及外源性转化生长因子TGF-β1(transforming growth factor-1)诱导BMSCs向肌成纤维细胞定向分化的可能性,为放射性诱导的颌骨纤维化提供体外理论依据。方法取大鼠股骨的骨髓,全髓培养法分离培养骨髓间充质干细胞;培养至第3代时分别用+/-TGF-β1的2组诱导培养基定向分化诱导;诱导2周后显微镜下观察细胞形态变化,Real-time PCR检测细胞的基因表达情况;细胞爬片行免疫组化鉴定α-SMA及胶原纤维的表达。结果大鼠股骨骨髓组织中可分离出骨髓间充质干细胞,体外生长形态类似成纤维细胞,呈梭型、纺锤型,定向诱导后TGF-β1组细胞可表现出肌成纤维细胞的特性(α-SMA+),同时伴有COLⅠ、COLⅢ、α-SMA基因的高表达。结论从大鼠股骨骨髓组织中可分离出骨髓间充质干细胞,TGF-β1能定向诱导BMSCs向肌成纤维细胞方向分化,为放射诱导的颌骨纤维萎缩机制提供了体外理论依据。
[Abstract]:Objective to investigate the isolation, culture and amplification of BMSCs (bone marrow mesenchymal stem cells) in rat bone marrow mesenchymal stem cells (MSCs), and the possibility of inducing the directional differentiation of BMSCs myofibroblast induced by exogenous transforming growth factor TGF- beta 1 (transforming growth factor-1), and to provide a theoretical basis for radioactive induced maxillofacial fibrosis. Methods the bone marrow of the rat femur was taken and the bone marrow mesenchymal stem cells were isolated and cultured in the full marrow culture method. When cultured to third generations, 2 groups of +/-TGF- beta 1 were induced to induce differentiation, and the morphological changes of the cells were observed under the microscope after 2 weeks. The gene expression of the cells was detected by Real-time PCR, and the cell crawling film was used to identify the alpha -SMA by immunohistochemistry. Results the bone marrow mesenchymal stem cells can be separated from the bone marrow tissue of the rat femur. The growth morphology is similar to the fibroblast, spindle type and spindle type. After directed induction, TGF- beta 1 groups can show the characteristics of myofibroblast (alpha -SMA+), accompanied by the high expression of COL I, COL III, and alpha -SMA gene. Bone marrow mesenchymal stem cells can be separated from the bone marrow tissue of the rat femur. TGF- beta 1 can induce BMSCs to differentiate into myofibroblast direction, which provides a theoretical basis for the mechanism of radiation-induced atrophy of the jaws.

【作者单位】：上海交通大学医学院附属第九人民医院口腔医学院口腔颌面头颈肿瘤科上海市口腔医学重点实验室;上海交通大学医学院附属第九人民医院口腔医学院口腔放疗科;
【基金】：国家自然科学基金(81271112) 上海市教委“曙光计划”(10SG19)
【分类号】：R329.26

【共引文献】