绿色荧光蛋白标记成人骨髓间充质干细胞的超微结构特征

背景：研究表明,绿色荧光蛋白标记的骨髓间充质干细胞免疫学表型、细胞周期、分化潜能等均未发生明显改变。目的：进一步观察绿色荧光蛋白标记对骨髓间充质干细胞超微结构的影响。方法：分离培养成人骨髓间充质干细胞,抗CD34、抗CD105免疫细胞化学染色鉴定,选第3代细胞进行绿色荧光蛋白标记,超薄切片后用透射电镜观察细胞超微结构,以未经绿色荧光蛋白标记骨髓间充质干细胞为对照。结果与结论：骨髓间充质干细胞表达CD105,不表达CD34。绿色荧光蛋白标记后,骨髓间充质干细胞细胞质发出绿色荧光,绿色荧光主要集中于细胞核周围的胞浆内,远离细胞核的胞浆内荧光强度逐渐减弱。相对于未标记的骨髓间充质细胞,经绿色荧光蛋白标记后细胞粗面内质网、高尔基体等细胞器较多,而线粒体相对较少;另外,脂滴和＂空泡＂样结构也多一些。结果证实绿色荧光蛋白对骨髓间充质干细胞性状的影响较为局限,是一种较为理想的细胞标记示踪剂。

Ultrastructural characteristics of adult bone marrow mesenchymal stem cells labeled with green fluorescent protein

BACKGROUND：Studies have suggested that no significant changes were determined in immunological phenotype,cell cycle and differentiation potential of bone marrow mesenchymal stem cells（BMSCs） labeled with green fluorescent protein（GFP）.OBJECTIVE：To further study effects of GFP on ultrastructure of BMSCs.METHODS：BMSCs were isolated from adult marrow and cultured in vitro.BMSCs were stained in immunocytochemistry with antibody of anti-CD34 and anti-CD105 to identify.The third generation of BMSCs were labeled with GFP.Ultrathin sections of BMSCs labeled with GFP were observed for ultrastructure under the transmission electron microscope,and normal BMSCs without GFP served as controls.RESULTS AND CONCLUSION：BMSCs expressed CD105 and not CD34.They labeled with GFP had green fluorescence in the cytoplasm.The fluorescent strength became weak gradually in the cytoplasm away from nuclei.BMSCs labeled GFP had many rough endoplasm reticulum and Golgi body,less mitochondrion than normal BMSCs.In addition,lipid droplet and ＂vacuole＂-like structure became more.Results verified that effects of GFP on BMSCs were limited,and GFP was an ideal tracing reagent.