Expression data of mESCs differentiated into Paraxial mesoderm

Stem cell-derived tissues have wide potential for modelling developmental and pathological processes as well as cell-based therapy. However, it has proven difficult to generate several key cell types in vitro, including skeletal muscle. In vertebrates, skeletal muscles derive during embryogenesis from the presomitic mesoderm (PSM). Using PSM development as a guide, we establish conditions for the differentiation of monolayer cultures of mouse embryonic stem (ES) cells into PSM-like cells without the introduction of transgenes or cell sorting. We differentiated mouse ESCs in serum-free medium supplemented with Rspo3 ( or as an alternative with Chir 9902) and the Bmp inhibitor LDN193189. In vivo, the PSM cells are first expressing Msgn1 (posterior PSM marker) and then mature to express Pax3 (anterior PSM marker). After 4 days of differentiation of mESCs, Msgn1-positive cells were FACS-sorted and their transcriptome analyzed. After 6 days of differentiation, Pax3-positive cells were sorted and their transcriptome analyzed. Mouse ESCs differentiated for 0, 4 and 6 days in serum-free medium containing a Wnt activator, a BMP inhibitor and DMSO, to study paraxial mesoderm in vitro

干细胞衍生组织在发育与病理过程建模以及细胞治疗领域具备广阔应用前景。然而，体外诱导生成多种关键细胞类型始终存在技术瓶颈，其中便包括骨骼肌。在脊椎动物中，骨骼肌在胚胎发生过程中起源于体节前中胚层（presomitic mesoderm, PSM）。以体节前中胚层的发育进程为指导，我们建立了无需引入转基因或进行细胞分选的小鼠胚胎干细胞（embryonic stem cells, ES细胞）单层培养物向体节前中胚层样细胞分化的培养体系。我们在添加Rspo3（或替代方案为Chir 9902）与骨形态发生蛋白（bone morphogenetic protein, BMP）抑制剂LDN193189的无血清培养基中对小鼠胚胎干细胞进行诱导分化。体内研究表明，体节前中胚层细胞首先表达Msgn1（体节前中胚层后部标志物），随后成熟并表达Pax3（体节前中胚层前部标志物）。在小鼠胚胎干细胞诱导分化4天后，我们通过荧光激活细胞分选（fluorescence-activated cell sorting, FACS）分离得到Msgn1阳性细胞并对其转录组进行分析；分化6天后，则分离得到Pax3阳性细胞并开展转录组分析。本研究同时设置对照实验组：在添加Wnt激活剂、BMP抑制剂与二甲基亚砜（dimethyl sulfoxide, DMSO）的无血清培养基中，将小鼠胚胎干细胞分别诱导分化0天、4天及6天，以体外研究轴旁中胚层的相关特性。