脯氨酸羟化酶抑制剂DMOG动员的大鼠外周血间充质干细胞的多向分化潜能研究

发布时间：2018-09-18 10:40

【摘要】：目的:探讨脯氨酸羟化酶抑制剂—二甲基乙二酰基甘氨酸(DMOG)动员后大鼠外周血间充质干细胞(PB-MSCs)的多向分化潜能。方法:SD大鼠腹腔注射DMOG 40 mg·kg-1,分离扩增PB-MSCs,流式细胞术检测其表面抗原;诱导分化培养法研究PB-MSCs成骨、成软骨、成脂、成神经分化潜能。结果:PB-MSCs表面阴性表达造血系标记CD34(2.16±1.60)%、CD45(0.82±0.55)%,阳性表达CD90(98.66±0.77)%;成骨分化检测有钙结节出现,硝酸银染色发现细胞成黑色颗粒状;成软骨分化检测发现细胞核偏移,甲苯胺蓝染色细胞外基质成紫色,核成蓝色;成脂分化检测发现细胞中出现大量"脂滴",油红O染色成红色;成神经检测中,细胞出现轴突,免疫荧光蛋白检测巢蛋白(Nestin)表达阳性,胶质纤维酸性蛋白(GFAP)表达阴性。结论:DMOG动员后培养的外周血细胞为MSCs,且具有成骨、成软骨、成脂、成神经分化的潜能。
[Abstract]:Aim: to investigate the multidirectional differentiation potential of rat peripheral blood mesenchymal stem cells (PB-MSCs) mobilized by dimethyl acetylglycine (DMOG), a proline hydroxylase inhibitor. Methods DMOG 40 mg kg-1, was injected intraperitoneally to detect the surface antigen of SD rats, and PB-MSCs osteogenesis, cartilage formation, adipogenesis and neural differentiation potential were studied by induced differentiation culture. Results the hematopoietic marker CD34 was (2.16 卤1.60) and CD45 was positive (0.82 卤0.55), and the positive expression of CD90 was (98.66 卤0.77). Toluidine blue stained the extracellular matrix purple and the nucleus blue; the adipogenic differentiation test revealed a large number of "lipid droplets" in the cells, and the oil red O staining became red; in the neurogenesis test, the axons appeared in the cells. Positive expression of nestin (Nestin) and negative expression of glial fibrillary acidic protein (GFAP) (GFAP) were detected by immunofluorescence. Conclusion the peripheral blood cells cultured after the mobilization of MSCs, have the potential of osteogenesis, cartilage formation, lipid formation and neural differentiation.
【作者单位】：浙江中医药大学生命科学学院;浙江中医药大学基础医学院;
【基金】：国家自然科学基金(81270566) 浙江省自然科学基金(LY15C100001)
【分类号】：R965

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