Vural Rengim, Çelik Dicle, Peker Ersin Berkay, Köni Ekin, Kulaç Ayşe Aydanur, Tokcaer Keskin Zeynep
Department of Molecular and Translational Biomedicine, Graduate School of Natural and Applied Sciences, Acıbadem Mehmet Ali Aydınlar University, İstanbul, Turkey.
Acıbadem Labcell, İstanbul, Turkey.
Turk J Biol. 2022 Jul 14;46(5):375-387. doi: 10.55730/1300-0152.2624. eCollection 2022.
The effects of () 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 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 genotype in hBM-MSCs.
本研究调查了()C677T基因多态性对人骨髓间充质干细胞(hBM-MSCs)活力、形态、生理及分化能力的影响。为此,在获得伦理委员会许可并告知供体的情况下,获取了该基因野生型(WT,C/C)、杂合子(HTZ,C/T)和纯合子(HMZ,T/T)的原代hBM-MSCs。使用RFLP和桑格测序方法从细胞分离的基因组DNA中检测突变,通过CFU-F试验研究集落形成特性,通过MTT试验研究增殖差异。诱导脂肪生成、成骨和成软骨分化以研究其分化潜能的变化,并使用Graphpad Prism软件进行单因素方差分析对结果进行统计学分析。共筛选了13名供体,细胞的hBM-MSC标志物和体外形态无差异。CFU-F试验结果显示WT与HTZ之间存在显著差异,但24小时和48小时后的MTT试验无显著差异。分化试验结果表明,HMZ细胞的脂肪生成分化明显多于WT细胞。成骨和成软骨分化结果无统计学显著差异。这些实验结果表明,hBM-MSCs的脂肪生成分化受该基因基因型的影响。