Department of Molecular Cell Biology, Life Sciences Center, Institute of Biochemistry, Vilnius University, Saulėtekio Av. 7, 10257, Vilnius, Lithuania.
Faculty of Physics, Laser Research Center, Vilnius University, Saulėtekio Av. 10, 10223, Vilnius, Lithuania.
Sci Rep. 2024 Jul 23;14(1):16966. doi: 10.1038/s41598-024-68108-0.
Stem cells' differentiation toward cardiac lineage is a complex process dependent on various alterations in molecular basis and regulation pathways. The aim of the study is to show that endometrium-derived stromal cells - menstrual, endometrial and endometriotic, could be an attractive source for examination of the mechanisms underlying cardiomyogenesis. After treatment with Decitabine, Angiotensin II and TGF-β1, cells demonstrated morphological dedifferentiation into early cardiomyocyte-like cells and expressed CD36, CD106, CD172a typically used to sort for human pluripotent stem cell-derived cardiomyocytes. RT-qPCR revealed changed cells' genetic profiles, as majority of cardiac lineage differentiation related genes and cardiac ion channels (calcium, sodium, potassium) coding genes were upregulated after 6 and 13 days of exposure. Additionally, analysis of expression of various signaling proteins (FOXO1, PDGFB, TGFBR1, mTOR, VEGFA, WNT4, Notch1) coding genes showed differences between cell cultures as they seem to employ distinct signaling pathways through differentiation initiation. Early stages of differentiation had biggest impact on cardiomyogenesis related proteins (Nkx-2.5, EZH2, FOXO3a, H3K9Ac) levels, as we noticed after conducting Western blot and as expected, early cardiac transcription factor Nkx-2.5 was highly expressed and localized in nucleus of differentiating cells. These findings led us to assess endometrium origin stromal cells' potential to differentiate towards cardiomyogenic lineage and better understand the regulation of complex differentiation processes in ex vivo model systems.
干细胞向心脏谱系的分化是一个复杂的过程，依赖于分子基础和调节途径的各种改变。本研究的目的是表明子宫内膜基质细胞——月经、子宫内膜和子宫内膜异位症——可以成为研究心脏发生机制的有吸引力的来源。在用地西他滨、血管紧张素 II 和 TGF-β1 处理后，细胞表现出形态去分化为早期心肌样细胞，并表达通常用于分选人多能干细胞衍生的心肌细胞的 CD36、CD106、CD172a。RT-qPCR 揭示了细胞遗传特征的变化，因为大多数心脏谱系分化相关基因和心脏离子通道（钙、钠、钾）编码基因在暴露 6 和 13 天后上调。此外，分析各种信号蛋白（FOXO1、PDGFB、TGFBR1、mTOR、VEGFA、WNT4、Notch1）编码基因的表达表明细胞培养之间存在差异，因为它们似乎通过分化起始采用不同的信号通路。分化的早期阶段对心肌发生相关蛋白（Nkx-2.5、EZH2、FOXO3a、H3K9Ac）水平有最大的影响，这是我们在进行 Western blot 后注意到的，并且正如预期的那样，早期心脏转录因子 Nkx-2.5 高度表达并定位于分化细胞的核内。这些发现使我们能够评估子宫内膜来源基质细胞向心肌谱系分化的潜力，并更好地理解复杂分化过程在体外模型系统中的调控。
