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鸡胚胎干细胞能量代谢的特点

, PP. 938-951

Keywords: chES细胞,能量代谢,糖原,细胞命运,6-磷酸葡萄糖

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Abstract:

细胞能量代谢与细胞生命活动紧密相关,但目前对于鸡胚胎干细胞能量代谢特点的研究较少.本研究利用前期建立的chES细胞模型,通过电子显微镜发现未分化的chES细胞胞质内储存大量糖原颗粒和卵黄颗粒.逆转录-聚合酶链反应对能量代谢途径中的主要限速酶的基因表达分析显示,未分化的chES细胞不表达丙酮酸羧化酶和磷酸烯醇式丙酮酸羧激酶,与鸡胚胎成纤维细胞相比低表达葡萄糖转运蛋白1和磷酸果糖激酶,高表达己糖激酶1和糖原合成酶;分化中的chES细胞与成年母鸡肝脏相比少量表达PCX和PCK2,与CEF细胞相比高表达GLUT1,HK1,PFK,而GYS的表达没有显著性差异.过碘酸希夫氏染色显示,细胞培养过程中未分化chES细胞中储存的糖原颗粒未见减少,而分化中的chES细胞的糖原颗粒逐渐减少.此外,通过赫斯特荧光染料33342和碘化丙啶双染色发现,大量未分化的chES细胞在体外培养过程中死亡,而添加外源性6-磷酸葡萄糖显著减少细胞死亡.上述结果说明,未分化chES细胞主要以糖原合成为主,无糖异生代谢,而分化中的chES细胞以糖酵解代谢为主.未分化的chES细胞以大量消耗代谢中间产物G6P为代价进行糖原合成,造成细胞内源性G6P相对不足引起细胞死亡.因此,细胞内糖原的含量反映了chES细胞的分化程度.chES细胞在体外培养时可通过添加G6P来提高生存率.

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