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2型糖尿病血清对脐带来源间充质干细胞行为的影响

Effect of type 2 diabetic serum on the behavior of Wharton's jelly-derived mesenchymal stem cells .

作者信息

Ali Fatima, Aziz Fehmina, Wajid Nadia

机构信息

Institute of Molecular Biology and Biotechnology (IMBB) & Centre for Research In Molecular Medicine (CRIMM), The University of Lahore, Raiwind Road, Lahore, Pakistan.

出版信息

Chronic Dis Transl Med. 2017 Apr 17;3(2):105-111. doi: 10.1016/j.cdtm.2017.02.006. eCollection 2017 Jun 25.

Abstract

OBJECTIVE

Wharton jelly-derived mesenchymal stem cells (WJMSCs) exhibit multilineage differentiation potential and can be used to treat multiple organs. However, diabetes affects the repair capability of MSCs. The aim of this study was to evaluate the effect of diabetic patient-derived serum on WJMSC behavior.

METHODS

WJMSCs at passage 3 were treated with serum derived from type 2 diabetic patients. WJMSCs were characterized for surface markers expression by using immunocytochemistry technique. The effects on cell viability, proliferation, cell death rate, and vascular endothelial growth factor level were assessed by crystal violet staining, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, lactate dehydrogenase assay, and enzyme-linked immuno-sorbent assay, respectively. Oxidative stress was assessed by the estimation of free radical species malondialdehyde (MDA) and enzymes glutathione (GSH), catalase, and superoxide dismutase (SOD).

RESULTS

WJMSCs isolated in this study were positive for CD29, CD49, CD73, CD90, CD105, and SSEA4 and negative for CD45 and CD34. Under diabetic stress conditions, WJMSCs showed low viability and high lactate dehydrogenase release. A low level of vascular endothelial growth factor was also observed after diabetic serum treatment. Antioxidant level was also lower in diabetic serum-treated WJMSCs compared to in normal serum-treated WJMSCs.

CONCLUSION

The results of the present study suggest that pre-treatment of WJMSCs with type 2 diabetic serum decreases the survival of WJMSCs. The findings of this study provide insight into diabetes-induced harmful effects on WJMSCs.

摘要

目的

脐带来源的间充质干细胞(WJMSCs)具有多向分化潜能,可用于治疗多个器官。然而,糖尿病会影响间充质干细胞的修复能力。本研究旨在评估糖尿病患者来源的血清对WJMSC行为的影响。

方法

用2型糖尿病患者来源的血清处理第3代WJMSCs。采用免疫细胞化学技术对WJMSCs的表面标志物表达进行鉴定。分别通过结晶紫染色、3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法、乳酸脱氢酶法和酶联免疫吸附测定法评估对细胞活力、增殖、细胞死亡率和血管内皮生长因子水平的影响。通过估计自由基物质丙二醛(MDA)和酶谷胱甘肽(GSH)、过氧化氢酶和超氧化物歧化酶(SOD)来评估氧化应激。

结果

本研究分离的WJMSCs对CD29、CD49、CD73、CD90、CD105和SSEA4呈阳性,对CD45和CD34呈阴性。在糖尿病应激条件下,WJMSCs显示出低活力和高乳酸脱氢酶释放。糖尿病血清处理后还观察到血管内皮生长因子水平较低。与正常血清处理的WJMSCs相比,糖尿病血清处理的WJMSCs中的抗氧化水平也较低。

结论

本研究结果表明,用2型糖尿病血清预处理WJMSCs会降低WJMSCs的存活率。本研究结果为糖尿病对WJMSCs的有害影响提供了见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1bde/5627692/fe49176b42d4/gr1.jpg

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