Asgharian Razieh, Hashemi Afrooz, Javeri Arash, Fakhr Taha Masoumeh
Department of Stem Cells and Regenerative Medicine, Institute for Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
Department of Biology, Islamic Azad University, Gorgan Branch, Gorgan, Iran.
Iran J Biotechnol. 2024 Apr 1;22(2):e3863. doi: 10.30498/ijb.2024.435096.3863. eCollection 2024 Apr.
DNA methylation plays important roles in regulating various biological processes, including self-renewal, differentiation and regenerative capacity of stem cells. Previous studies have demonstrated that lineage-specific differentiation of mesenchymal stem cells can be promoted using nontoxic chromatin-modifying drugs.
Here we evaluated the impact of RG108, a known DNA methyltransferase inhibitor, on the expression of pluripotency genes in human adipose tissue-derived stem cells (hADSCs) and their proliferation and differentiation.
Human ADSCs were isolated by collagenase treatment and characterized. Then, ADSCs were treated with 5 µM RG108 for four days. The control and RG108-treated cells were analyzed for the cell cycle progression, apoptosis and the expression of pluripotency genes. Also, ADSCs were cultured in adipogenic and osteogenic differentiation media for three weeks and were assessed by Oil Red O and Alizarin Red S staining and qPCR analysis.
We showed that RG108 treatment increased proliferation of hADSCs and upregulated the expression of pluripotency-related genes. Additionally, RG108 had a positive impact on the differentiation capability of ADSCs. This was evident through elevated levels of Oil Red O staining in the RG108 treatment group. Also, qPCR analysis showed the upregulation of some adipogenic and osteogenic markers by RG108.
These findings indicate that pretreatment with RG108 improves the differentiation potential of ADSCs, probably making these cells more beneficial for cell therapy applications.
DNA甲基化在调节各种生物学过程中发挥重要作用，包括干细胞的自我更新、分化和再生能力。先前的研究表明，使用无毒的染色质修饰药物可以促进间充质干细胞的谱系特异性分化。
在此，我们评估了已知的DNA甲基转移酶抑制剂RG108对人脂肪组织来源干细胞（hADSCs）多能性基因表达及其增殖和分化的影响。
通过胶原酶处理分离并鉴定人ADSCs。然后，将ADSCs用5μM RG108处理4天。分析对照细胞和经RG108处理的细胞的细胞周期进程、凋亡情况以及多能性基因的表达。此外，将ADSCs在成脂和成骨分化培养基中培养3周，并通过油红O和茜素红S染色以及qPCR分析进行评估。
我们发现RG108处理可增加hADSCs的增殖并上调多能性相关基因的表达。此外，RG108对ADSCs的分化能力有积极影响。这在RG108处理组中油红O染色水平升高上得以体现。而且，qPCR分析显示RG108可上调一些成脂和成骨标志物的表达。
这些发现表明，用RG108预处理可提高ADSCs的分化潜能，可能使这些细胞在细胞治疗应用中更具优势。
