Department of Embryo Transfer Research, Gyeongbuk Livestock Research Institute, Yeongju, Korea.
Core Protein Resources Center, DGIST, Daegu, Korea.
Cell Reprogram. 2020 Oct;22(5):244-253. doi: 10.1089/cell.2020.0006. Epub 2020 Sep 16.
Mouse embryonic stem cells (mESCs) go through self-renewal in the existence of the cytokine leukemia inhibitory factor (LIF). LIF is added to the mouse stem cells culture medium, and its removal results in fast differentiation. Dimethyl sulfoxide (DMSO) is one of the most used solvents in drug test. We exposed 4-day mESC cultures to different concentrations of DMSO (0.1%, 0.5%, 1.0%, and 2.0%) to identify the safest dose exhibiting efficacy as a solvent. mESCs grown under general pluripotency conditions in the absence of LIF were treated with DMSO. In addition, as a control for differentiation, mESCs were grown in the absence of LIF. DMSO upregulated the mRNA expression level of pluripotency markers. Moreover, DMSO reduced the mRNA expression levels of ectodermal marker (β-tubulin3), mesodermal marker (Hand1), and endodermal markers (Foxa2 and Sox17) in mESCs. These results indicate that DMSO treatment enhances the pluripotency and disrupts the differentiation of mESCs. We also show that members of the Tet oncogene family are critical to inhibiting the differentiation and methylation of mESCs. DMSO is appropriate to sustain the pluripotency of mESCs in the absence of LIF, and that mESCs can be sustained in an undifferentiated state using DMSO. Therefore, DMSO may, in part, function as a substitute for LIF.
小鼠胚胎干细胞（mESCs）在细胞因子白血病抑制因子（LIF）的存在下进行自我更新。将 LIF 添加到小鼠干细胞培养基中，去除后会导致快速分化。二甲基亚砜（DMSO）是药物测试中最常用的溶剂之一。我们将 4 天的 mESC 培养物暴露于不同浓度的 DMSO（0.1%、0.5%、1.0%和 2.0%）中，以确定作为溶剂表现出疗效的最安全剂量。在没有 LIF 的情况下，在一般多能性条件下生长的 mESCs 用 DMSO 处理。此外，作为分化对照，mESCs 在没有 LIF 的情况下生长。DMSO 上调了多能性标志物的 mRNA 表达水平。此外，DMSO 降低了 mESCs 中外胚层标志物（β-微管蛋白 3）、中胚层标志物（Hand1）和内胚层标志物（Foxa2 和 Sox17）的 mRNA 表达水平。这些结果表明 DMSO 处理增强了 mESCs 的多能性并破坏了其分化。我们还表明，Tet 癌基因家族成员对于抑制 mESCs 的分化和甲基化至关重要。DMSO 适合在没有 LIF 的情况下维持 mESCs 的多能性，并且可以使用 DMSO 将 mESCs 维持在未分化状态。因此，DMSO 可能部分起到 LIF 的替代作用。
