1-磷酸鞘氨醇促进间充质干细胞向心肌分化

发布时间：2018-03-24 10:50

本文选题：脐带间充质干细胞　切入点：脂肪间充质干细胞　出处：《生物工程学报》2013年11期

【摘要】：为研究1-磷酸鞘氨醇(Sphingosine-1-phosphate,S1P)对脐带间充质干细胞(Umbilical cord mesenchymal stem cells,UC-MSCs)和脂肪间充质干细胞(Adipose derived mesenchymal stem cells,AD-MSCs)向心肌分化的影响,探索其适宜的作用时间和浓度,将UC-MSCs和AD-MSCs接种到培养板,用添加不同浓度S1P的心肌细胞培养液诱导两种干细胞向心肌分化,诱导时间分为7 d、14 d和28 d。采用免疫荧光染色检测心肌特异性蛋白,α-肌动蛋白(α-actin)、缝隙连接蛋白(Connexin-43)以及肌球蛋白重链(MYH-6)的表达,并通过共聚焦显微镜和荧光显微镜进行观察;采用MTT分析细胞的活性;膜片钳检测分化细胞的钙瞬变(此为心肌细胞的功能性指标)。结果表明,S1P与心肌细胞培养液协同作用,能够促进UC-MSCs和AD-MSCs向心肌细胞的分化。并且,随着S1P浓度的增加,促分化作用增强,但细胞活性降低。S1P在心肌细胞培养液中的适宜作用时间为14 d,适宜作用浓度为0.5μmol/L。而且联合心肌细胞培养液可以使UC-MSCs和AD-MSCs的心肌分化细胞产生钙瞬变,具有类似心肌细胞的功能性。S1P能够与心肌细胞培养液协同作用,促进UC-MSCs和AD-MSCs的心肌功能性分化。
[Abstract]:In order to study the effects of Sphingosine-1-phosphate S1P on myocardial differentiation of umbilical cord mesenchymal stem cells (Umbilical cord mesenchymal stem stem cells) and adipose mesenchymal stem cells (adipose derived mesenchymal stem cells AD-MSCs), and to explore the appropriate time and concentration of Sphingosine-1-phosphateanine, UC-MSCs and AD-MSCs were inoculated into the culture plate. Two kinds of stem cells were induced to differentiate into myocardium by adding different concentrations of S1P into cardiomyocyte culture medium. The induction time was divided into 7 days, 14 days and 28 days. The expression of myocardial specific protein, 伪 -actin (伪 -actin), gap junction protein (gap junction protein) and myosin heavy chain myosin (MYH-6) were detected by immunofluorescence staining, and were observed by confocal microscope and fluorescence microscope. MTT was used to analyze cell activity, patch clamp was used to detect calcium transient of differentiated cells (this is the functional index of cardiomyocytes). The results showed that S1P could promote the differentiation of UC-MSCs and AD-MSCs into cardiomyocytes by synergistic action with cardiomyocyte culture medium. With the increase of S1P concentration, the effect of promoting differentiation was enhanced. However, the suitable time for the decrease of S1P activity in cardiomyocyte culture medium was 14 days and the optimal concentration was 0.5 渭 mol / L. moreover, combined with cardiomyocyte culture medium, calcium transient was produced in myocardial differentiation cells of UC-MSCs and AD-MSCs. S1P, which is similar to cardiomyocytes, can promote myocardial functional differentiation of UC-MSCs and AD-MSCs by synergistic action with cardiomyocyte culture medium.
【作者单位】：大连理工大学化工学院干细胞与组织工程研发中心;大连理工大学盘锦校区生命与医药学院;
【基金】：国家自然科学基金(No.31170945)资助~~
【分类号】：R329

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