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

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。

Objective: To explore the multi-directional differentiation of rat peripheral blood mesenchymal stem cells(PB-MSCs)mobilized by proline hydroxylase inhibitor,dimethyloxaloylglycine(DMOG).Methods: SD rats were injected intraperitoneally with DMOG 40 mg·kg^-1, then the PB-MSCs were harvested and amplified in vitro,and the surface antigens of PB-MSCs were measured by flow cytometry. The capability of PB-MSCs multidirectional differentiation, including osteogenesis, chondrogenesis, adipogenesis and neural differentiation, were investigated by cell-culture inoculation test in vitro.Results: PB-MSCs were negative for CD34(2. 16±1. 60)%, CD45(0. 82±0. 55)%,and positive for CD90(98. 66±0. 77)%. The osteogenic differentiation assay detected the formation of calcium nodules; Silver nitrate staining found that the cells were blackish granular; Cartilage differentiation assay detected the nuclear migration,extracellular matrix were dyed purple with toluidine blue,the nucleus into blue. A large number of "lipid droplets" were observed after adipogenic differentiation,and cells were dyed red with oil red O. The axons appeared in the process of neural differentiation,then the expression of Nestin and glial fibrillary acidic peotein(GFAP)were respectively positive and negative.Conclusion: Peripheral blood cells mobilized by DMOG were proved to be MSCs possess potential capability of chondrogenesis, osteogenesis, adipogenesis and neural differentiation.