Maioli Margherita, Contini Giovanni, Santaniello Sara, Bandiera Pasquale, Pigliaru Gianfranco, Sanna Raimonda, Rinaldi Salvatore, Delitala Alessandro P, Montella Andrea, Bagella Luigi, Ventura Carlo
Department of Biomedical sciences, University of Sassari, Sassari, Italy ; Laboratory of Molecular Biology and Stem Cell Engineering, National Institute of Biostructures and Biosystems, Bologna, Italy ; Department of Regenerative Medicine, Rinaldi Fontani Institute, Florence, Italy.
Drug Des Devel Ther. 2013 Sep 27;7:1063-73. doi: 10.2147/DDDT.S44706. eCollection 2013.
Mouse embryonic stem cells were previously observed along with mesenchymal stem cells from different sources, after being treated with a mixed ester of hyaluronan with butyric and retinoic acids, to show a significant increase in the yield of cardiogenic and vascular differentiated elements. The aim of the present study was to determine if stem cells derived from primitive fetal cells present in human amniotic fluid (hAFSCs) and cultured in the presence of a mixture of hyaluronic (HA), butyric (BU), and retinoic (RA) acids show a higher yield of differentiation toward the cardiovascular phenotype as compared with untreated cells. During the differentiation process elicited by exposure to HA + BU + RA, genes controlling pluripotency and plasticity of stem cells, such as Sox2, Nanog, and Oct4, were significantly downregulated at the transcriptional level. At this point, a significant increase in expression of genes controlling the appearance of cardiogenic and vascular lineages in HA + BU + RA-treated cells was observed. The protein expression levels typical of cardiac and vascular phenotypes, evaluated by Western blotting, immunofluorescence, and flow cytometry, were higher in hAFSCs cultured in the presence of HA + BU + RA, as compared with untreated control cells. Appearance of the cardiac phenotype was further inferred by ultrastructural analysis using transmission and scanning electron microscopy. These results demonstrate that a mixture of HA + BU + RA significantly increased the yield of elements committed toward cardiac and vascular phenotypes, confirming what we have previously observed in other cellular types.
先前观察到，在用透明质酸与丁酸和视黄酸的混合酯处理后，小鼠胚胎干细胞与来自不同来源的间充质干细胞一起，向心脏和血管分化成分的产量显著增加。本研究的目的是确定源自人羊水（hAFSCs）中原始胎儿细胞并在透明质酸（HA）、丁酸（BU）和视黄酸（RA）混合物存在下培养的干细胞，与未处理的细胞相比，是否向心血管表型的分化产量更高。在暴露于HA + BU + RA引发的分化过程中，控制干细胞多能性和可塑性的基因，如Sox2、Nanog和Oct4，在转录水平上显著下调。此时，观察到在HA + BU + RA处理的细胞中，控制心脏和血管谱系出现的基因表达显著增加。通过蛋白质印迹、免疫荧光和流式细胞术评估，与未处理的对照细胞相比，在HA + BU + RA存在下培养的hAFSCs中，心脏和血管表型典型的蛋白质表达水平更高。使用透射和扫描电子显微镜进行的超微结构分析进一步推断出心脏表型的出现。这些结果表明，HA + BU + RA混合物显著提高了向心脏和血管表型分化的成分产量，证实了我们之前在其他细胞类型中观察到的结果。
