本文選題：人胚胎干細(xì)胞　切入點：Apelin　出處：《山東醫(yī)藥》2017年33期 　論文類型：期刊論文

【摘要】：目的探討Apelin13對人胚胎干細(xì)胞(hESCs)定向心肌細(xì)胞分化效率的影響。方法將hESCs單層培養(yǎng)后經(jīng)EDTA消化傳代,應(yīng)用明確的誘導(dǎo)分化培養(yǎng)基對觀察組(加Apelin13)和對照組(不加Apelin13)定向誘導(dǎo)hESCs向心肌細(xì)胞分化。于誘導(dǎo)第2天(中胚層階段)、第4天(心肌分化初始階段)、第7天(心肌分化末階段)作為時間節(jié)點,采用實時熒光定量PCR檢測兩組Brachury T、Mesp1、NKx2.5、APJ mRNA表達(dá)水平,采用Western blotting檢測兩組Brachury T、Mesp1、NKx2.5、APJ、Nanog、Sox2蛋白表達(dá);同時于誘導(dǎo)第7天采用細(xì)胞免疫熒光技術(shù)檢測兩組心肌細(xì)胞標(biāo)志物肌鈣蛋白T2(TNNT2)、α-輔肌蛋白(α-actinin)表達(dá)。結(jié)果誘導(dǎo)第7天觀察組hESCs定向心肌分化程度高于對照組,免疫熒光染色倒置顯微鏡下可清楚看到成熟心肌細(xì)胞的肌結(jié)及心肌細(xì)胞TNNT2、α-actinin表達(dá)。在3個誘導(dǎo)時間節(jié)點,觀察組Brachury T、Mesp1、NKx2.5、APJ mRNA及其蛋白表達(dá)均高于對照組(P均0.05),觀察組hESCs標(biāo)志蛋白Nanog、Sox2低于對照組(P均0.05)。結(jié)論 Apelin13可提高h(yuǎn)ESCs向心肌細(xì)胞的分化效率。
[Abstract]:Objective to investigate the effect of Apelin13 on the differentiation efficiency of human embryonic stem cells (hESCs) oriented cardiomyocytes. Methods hESCs monolayer was cultured and subcultured with EDTA. HESCs was induced to differentiate into cardiomyocytes in the observation group (plus Apelin 13) and the control group (without Apelin 13). On the second day of induction (mesoderm stage, day 4, myocardial differentiation initial stage, day 7), the differentiation of cardiomyocytes was induced on the specific differentiation medium of the observation group (plus Apelin 13) and the control group (without Apelin 13). End stage of myocardial differentiation) as a time node, Real time fluorescence quantitative PCR was used to detect the expression of Brachury Mesp1 nkx2.5APJ mRNA, and Western blotting was used to detect the expression of Brachury Tet Mesp1 NKx2.5APG Sox2 protein. At the same time, on the 7th day after induction, the expression of troponin T _ 2 and 偽 -actinin (偽 -actinin) was detected by cellular immunofluorescence technique. Results on the 7th day of induction, the degree of hESCs directional myocardial differentiation in the observation group was higher than that in the control group. The myocyte node and the expression of TNNT2, 偽 -actinin in the mature cardiomyocytes were clearly observed by immunofluorescence staining under inverted microscope, and the expression of TNNT2, 偽 -actinin was observed at three induced time nodes. The expression of mRNA and its protein in Brachury Tet Mesp1 NKx2.5 Brachury in the observation group was higher than that in the control group (P 0.05), and the hESCs marker protein Nanogan Sox2 in the observation group was lower than that in the control group (P 0.05). Conclusion Apelin13 can increase the differentiation efficiency of hESCs into cardiomyocytes.
【作者單位】： 安徽醫(yī)科大學(xué)海軍臨床學(xué)院;中國人民解放軍海軍總醫(yī)院;
【基金】：國家自然科學(xué)基金資助項目(81370238) 北京市自然科學(xué)基金資助項目(7142156)
【分類號】：R54

【相似文獻(xiàn)】

相關(guān)期刊論文 前2條

1 ;人胚胎干細(xì)胞生成紅細(xì)胞、輸血不再愁[J];畜牧獸醫(yī)科技信息;2008年10期

2 余國膺;基因工程制成的人胚胎干細(xì)胞具有電活動的心臟生出物與受體不活動心室細(xì)胞的功能整合[J];中國介入心臟病學(xué)雜志;2005年03期

相關(guān)博士學(xué)位論文 前1條

1 李卓;核糖體基因區(qū)打靶載體介導(dǎo)FVⅢ打靶人胚胎干細(xì)胞及其定向分化研究[D];中南大學(xué);2013年

相關(guān)碩士學(xué)位論文 前1條

1 魏晉;人胚胎干細(xì)胞向心室和心房肌細(xì)胞分化和富集的研究[D];西南大學(xué);2012年

，

本文編號：1598601