MTHFR C677T突变影响人骨髓间充质干细胞成脂分化能力。

Rengim Vural, Dicle Çelik, Ersin Berkay Peker, Ekin Köni, Ayşe Aydanur Kulaç, Zeynep Tokcaer Keskin
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摘要

研究了5,10-亚甲基四氢叶酸还原酶(MTHFR) C677T多态性对人骨髓间充质干细胞(hBM-MSCs)活力、形态、生理和分化能力的影响。为此,经伦理委员会许可并告知供者,获得了MTHFR基因野生型(WT, C/C)、杂合子(HTZ, C/T)和纯合子(HMZ, T/T)的原代hBM-MSCs。用RFLP和Sanger测序方法检测细胞基因组DNA的突变,用CFU-F试验研究定植特性,用MTT试验研究增殖差异。诱导成脂分化、成骨分化和成软骨分化,研究其分化潜能的变化,采用Graphpad Prism的单因素方差分析对结果进行统计学分析。总共筛选了13个供体,在hBM-MSC标记物和细胞的体外形态方面没有差异。CFU-F测试结果显示WT和HTZ有显著性差异,而24和48 h后MTT测试结果无显著性差异。分化测试结果发现HMZ细胞的成脂分化明显多于WT细胞。成骨和软骨分化结果没有统计学意义。通过这些实验,我们发现hBM-MSCs的成脂分化受到MTHFR基因型的影响。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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MTHFR C677T mutation affects adipogenic differentiation abilities of human bone marrow-derived mesenchymal stem cells.

The effects of 5,10-Methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism on human bone marrow-derived mesenchymal stem cells (hBM-MSCs) viability, morphology, physiology and differentiation capacity were investigated in this study. For this purpose, primary hBM-MSCs with wild type (WT, C/C), heterozygote (HTZ, C/T) and homozygote (HMZ, T/T) for the MTHFR gene were obtained with ethical committee permission and donor informed. Mutations were detected using RFLP and Sanger sequencing methods from genomic DNA isolated from cells, colonization properties were investigated by CFU-F test and proliferative differences were investigated by MTT test. Adipogenic, osteogenic, and chondrogenic differentiation were induced to study changes in their differentiation potentials, and the results were statistically analyzed using one-way ANOVA with Graphpad Prism. A total of 13 donors were screened and there were no differences in the hBM-MSC markers and in vitro morphologies of the cells. While there were significant differences between WT and HTZ as a result of the CFU-F test, there were no significant differences in the MTT test after 24 and 48 h. As a result of differentiation tests, it was found that adipogenic differentiation was significantly more in HMZ cells than WT cells. Osteogenic and chondrogenic differentiation results did not give statistically significant results. As a result of these experiments, adipogenic differentiation was found to be affected by the MTHFR genotype in hBM-MSCs.

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