Studies on lipid signals associated with differentiation propensity in mouse ES cells

了解调节多能干细胞的心脏分化的机制对于作为细胞治疗产品的心肌细胞的有效生成和扩增是必要的。我们分离了在心肌发生中具有不同特性的P19 CL 6细胞亚系，并使用转录组分析提取了24个与心肌发生相关的心肌发生相关候选基因（CMR）。通过RNAi敲低CMR基因揭示了18个CMR基因影响胚胎癌（EC）细胞中自发收缩或心脏标记基因的转录水平。我们还在小鼠胚胎干（ES）细胞中敲除CMR基因并诱导体外心脏分化。三个CMR基因，AW 551984，前列腺酰胺/前列腺素F合成酶和Cd 302，调节EC细胞和ES细胞的心脏分化。在ES细胞分化过程中，AW 551984的消耗减弱了早期心脏转录因子Nkx2.5的表达，而不影响多能性和早期中胚层标记基因的转录水平。在拟胚体形成过程中，Wnt/β-catenin信号转导的激活增强了ES细胞中AW 551984和Nkx2.5的表达。我们的研究结果表明，AW 551984是多能胚胎细胞心肌发生的一种新型调节剂，它将Wnt/β-catenin信号传导与Nkx2.5表达联系起来。

An understanding of the mechanism that regulates the cardiac differentiation of pluripotent stem cells is necessary for the effective generation and expansion of cardiomyocytes as cell therapy products. We isolated P19CL6 cell sublines that possess distinct properties in cardiomyogenesis and extracted 24 cardiomyogenesis-related candidate (CMR) genes correlated with cardiomyogenesis using a transcriptome analysis. Knockdown of the CMR genes by RNAi revealed that 18 CMR genes influence spontaneous contraction or transcript levels of cardiac marker genes in embryonal carcinoma (EC) cells. We also performed knockdown of the CMR genes in mouse embryonic stem (ES) cells and induced in vitro cardiac differentiation. Three CMR genes, AW551984, Prostamide/prostaglandin F synthase and Cd302, modulated the cardiac differentiation of both EC cells and ES cells. Depletion of AW551984 attenuated the expression of early cardiac transcription factor Nkx2.5 without affecting transcript levels of pluripotency and early mesoderm marker genes during ES cell differentiation. Activation of Wnt/β-catenin signaling enhanced the expression of both AW551984 and Nkx2.5 in ES cells during embryoid body formation. Our findings indicate that AW551984 is a novel regulator of cardiomyogenesis from pluripotent embryonic cells, which links Wnt/β-catenin signaling to Nkx2.5 expression.