【摘要】：目的:探討脯氨酸羥化酶抑制劑—二甲基乙二�；拾彼�(DMOG)動員后大鼠外周血間充質(zhì)干細胞(PB-MSCs)的多向分化潛能。方法:SD大鼠腹腔注射DMOG 40 mg·kg-1,分離擴增PB-MSCs,流式細胞術(shù)檢測其表面抗原;誘導(dǎo)分化培養(yǎng)法研究PB-MSCs成骨、成軟骨、成脂、成神經(jīng)分化潛能。結(jié)果:PB-MSCs表面陰性表達造血系標(biāo)記CD34(2.16±1.60)%、CD45(0.82±0.55)%,陽性表達CD90(98.66±0.77)%;成骨分化檢測有鈣結(jié)節(jié)出現(xiàn),硝酸銀染色發(fā)現(xiàn)細胞成黑色顆粒狀;成軟骨分化檢測發(fā)現(xiàn)細胞核偏移,甲苯胺藍染色細胞外基質(zhì)成紫色,核成藍色;成脂分化檢測發(fā)現(xiàn)細胞中出現(xiàn)大量"脂滴",油紅O染色成紅色;成神經(jīng)檢測中,細胞出現(xiàn)軸突,免疫熒光蛋白檢測巢蛋白(Nestin)表達陽性,膠質(zhì)纖維酸性蛋白(GFAP)表達陰性。結(jié)論:DMOG動員后培養(yǎng)的外周血細胞為MSCs,且具有成骨、成軟骨、成脂、成神經(jīng)分化的潛能。
[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 鹵1.60) and CD45 was positive (0.82 鹵0.55), and the positive expression of CD90 was (98.66 鹵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.
【作者單位】： 浙江中醫(yī)藥大學(xué)生命科學(xué)學(xué)院;浙江中醫(yī)藥大學(xué)基礎(chǔ)醫(yī)學(xué)院;
【基金】：國家自然科學(xué)基金(81270566) 浙江省自然科學(xué)基金(LY15C100001)
【分類號】：R965

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