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The effect of reduced glutathione on the chondrogenesis of human umbilical cord mesenchymal stem cells

机译:还原型谷胱甘肽对人脐带间充质干细胞软骨形成的影响

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It has been discussed whether reduced glutathione (GSH) could promote the chondrogenic differentiation ability of human umbilical cord mesenchymal stem cells (hUC-MSCs). hUC-MSCs were isolated from human umbilical cord and their specificity was identified, then induced into cartilage-like cells in chondrogenic induction medium with transforming growth factor beta 1 (TGF-β1), especially with GSH. The morphological change before and after induction was observed through inverted phase contrast microscope, Type II collagen (COL2-A1) and glycosaminoglycan (GAG) were analyzed qualitatively by Toluidine blue and immunofluorescence technique, respectively, the contents of COL2-A1 and GAG were estimated from the determination of hydroxyproline content and Alcian Blue method separately. The mRNA expressions of GAG and COL2-A1 were assayed by real-time fluorescence quantitative PCR. After continuously cultured for 21 days with GSH, Toluidine blue staining and immunofluorescence reaction were all positive in basic induction medium group (group B), basic induction medium +0.5% dimethylsulfoxide (DMSO) group (group BD) and basic induction medium +0.5% DMSO +500 μM GSH group (group BDG). Moreover, compared with group B and group BD, the contents of COL2-A1 and GAG in group BDG relatively increased and the mRNA expression level of COL2-A1 and GAG also comparatively increased (P
机译:已经讨论了还原型谷胱甘肽(GSH)是否可以促进人脐带间充质干细胞(hUC-MSC)的软骨分化能力。从人脐带中分离出hUC-MSCs,鉴定其特异性,然后在转化生长因子β1(TGF-β1),尤其是GSH的软骨诱导培养基中诱导成软骨样细胞。倒置相差显微镜观察诱导前后的形态变化,用甲苯胺蓝和免疫荧光技术分别定性分析II型胶原蛋白(COL2-A1)和糖胺聚糖(GAG),估算COL2-A1和GAG的含量。分别由羟脯氨酸含量的测定和阿尔辛蓝法测定。实时荧光定量PCR检测GAG和COL2-A1的mRNA表达。用GSH连续培养21天后,碱性诱导培养基组(B组),碱性诱导培养基+0.5 %二甲基亚砜(DMSO)组(BD组)和碱性诱导培养基+0.5的甲苯胺蓝染色和免疫荧光反应均为阳性。 %DMSO +500μMGSH组(BDG​​组)。此外,与B组和BD组相比,BDG组的COL2-A1和GAG含量相对增加,并且COL2-A1和GAG的mRNA表达水平也相对增加(P

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