骨髓间充质干细胞体外诱导分化为胰岛样细胞及其对坏死胰腺组织的修复

Induction of mesenchymal stem cells into islet-like cells in vitro and its repair on injured pancreas tissues

目的:探讨骨髓间充质干细胞(MSC)体外诱导转化为胰岛样细胞及其对坏死胰腺组织修复的可能性.方法:①取大鼠骨髓细胞,进行MSC培养纯化扩增,用bFGF、EGF、条件培养液、高糖培养基、B27、地塞米松、胰岛素等培养体系诱导MSC向胰岛细胞转化;双硫腙染色鉴定胰岛样细胞.②结扎大鼠胰腺组织及其血管6～7 h制作胰腺局部坏死模型.将分离、纯化、诱导的MSC和骨髓细胞用DAPI标记,分别移植到实验组(移植MSC)和对照组(移植骨髓单个核细胞).观察大鼠生存状态并于2周后取胰腺组织检测.结果:①体外扩增的MSC诱导转化后,细胞由初期的长梭形变成多角形或圆形并逐渐聚集成团.双硫腙染色大部分细胞团呈亮红色,初步表明MSC可诱导转化为胰岛样细胞.②实验组大鼠成活率达80%,坏死的胰腺组织被修复,变黑的胰腺组织基本恢复正常,胰腺组织中存在核呈兰色荧光的细胞;对照组的动物腹腔积水、粘连,2周内全部死亡.结论:大鼠骨髓MSC可在体外诱导转化为胰岛样细胞,并可修复损伤的胰腺组织.

Objective To study the induction of islet-like cell from bone marrow mesenchymal stem cells （MSCs） in vitro and its repair effects on injury pancreas tissues. Methods ①MSCs were cultivated and expanded in vitro and induced with cell factors like bFGF, EGF, high glucose DMEM media into islet-like cells. The induced cells were identified by dithizon staining. ② MSCs and bone marrow cells were marked with DAPI and implanted into experimental group （MSCs implantation） and control group （bone marrow mononuclear leukocyte implantation） . The survival rate and morphological changes were observed two weeks after implantation. Results ① After the amplified MSCs were induced in vitro, their forms changed from long spinde-shape to polygon or round and attached each other. Dithizon staining showed bright red color for most of induced cells. It indicated MSCs could be induced into islet-like cells. ②The survival rate of the experimental group was 80%. The injured pancreas tissues were repaired. Blue fluorescent was observed in repaired pancreas tissues. The animals in control group all died during two weeks. Conclusion MSCs can be induced into islet-like cells in vitro and repair the injured pancreas.