血小板衍生微粒增加脐带间充质干细胞中hTERT的表达

Maryam Samareh Salavati Pour, Fatemeh Hoseinpour Kasgari, A. Farsinejad, A. Fatemi, R. M. Khalilabadi
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引用次数: 3

摘要

引言:间充质干细胞由于其自我更新的潜力和分化为多种组织的能力而被广泛研究。然而,它们在体外的几个分裂的寿命有限,这改变了各种细胞特性,减少了它们的应用。目的:我们评估了血小板衍生微粒对hTERT基因表达的影响,hTERT是参与衰老和细胞寿命的主要因素之一。材料和方法:本研究采用脐带间充质干细胞。通过倒置显微镜评估细胞形态并使用流式细胞术鉴定相关表面标记物来表征细胞。通过以不同速度离心血小板袋制备血小板衍生微粒,并通过Bradford测定法测定其浓度。在30%汇合时,用50μg/mL的微粒处理MSCs 5天。然后提取RNA并合成cDNA。使用实时聚合酶链式反应(PCR)评估hTERT的定量表达。结果:从脐带组织中分离出成纤维细胞样细胞,并通过流式细胞术检测间充质表面标记物的存在来鉴定MSCs。实时PCR显示,与对照组相比,用血小板衍生的微粒处理hTERT的基因表达增加了三倍以上。结论:我们得出结论,血小板衍生微粒可能是一种潜在的安全有效的方法,可以增加骨髓间充质干细胞中hTERT基因的表达,最终延长其体外寿命。
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Platelet-Derived Microparticles Increase Expression of hTERT in Umbilical Cord Mesenchymal Stem Cells
Introduction: Mesenchymal stem cells (MSCs) are widely studied due to their self- renewal potential and capacity to differentiate into multiple tissues. However, they have a limited life span of several divisions in vitro, which alters various cellular characteristics and reduces their application. Aim: We evaluated the effect of platelet-derived microparticles on gene expression of hTERT, one of the main factors involved in aging and cell longevity. Materials and methods: Umbilical cord MSCs were used for this study. Cells were characterized by evaluating morphology via inverted microscope and identifying associated surface markers using flow cytometry. Platelet-derived microparticles were prepared by centrifuging platelet bags at varying speeds, and their concen- trations were determined by Bradford assay. At 30% confluency, MSCs were treated with 50 μg/mL of microparticles for five days. Then, RNA was extracted and cDNA was synthesized. Quantitative expression of hTERT was assessed using real-time polymerase chain reaction (PCR). Results: Fibroblast-like cells were isolated from umbilical cord tissue and MSCs were identified by the presence of mesenchymal surface markers via flow cytometry. Real- time PCR showed that gene expression of hTERT increased by more than three times when treated with platelet-derived microparticles, in comparison to expression of the control group. Conclusion: We concluded that platelet-derived microparticles may be a potentially safe and effective method to increase hTERT gene expression in MSCs, ultimately prolonging their life span in vitro. 
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