Metabolic properties of chicken embryonic stem cells
Metabolic properties of chicken embryonic stem cells
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鸡胚胎干细胞的代谢特性
DOI：
10.1007/s11427-010-4055-8
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发表时间：
2010-09-01
影响因子：
9.1
通讯作者：
Li Ning
中科院分区：
文献类型：
--
作者：
Li Jia;Zhang BaoLu;Li Ning
细胞能量代谢与细胞命运相关，但鸡胚胎干细胞（chES）的代谢特性尚不清楚。利用先前建立的chES细胞模型和电子显微镜（EM），我们发现未分化的chES细胞储存糖原。此外，与对照的原代鸡胚成纤维细胞（CEF）相比，未分化的chES细胞表达较低水平的葡萄糖转运蛋白1 （GLUT1）和磷酸果糖激酶（PFK） mrna，但表达较高水平的己糖激酶1 （HK1）和糖原合成酶（GYS） mrna，这表明chES细胞将葡萄糖通量导向糖原途径。此外，我们证明未分化的chES细胞阻断糖异生流出并阻碍葡萄糖-6-磷酸（G6P）从这一途径的积累，丙酮酸羧化酶（PCX）和线粒体磷酸烯醇丙酮酸羧化激酶（PCK2） mrna的水平几乎检测不到。此外，Hoechst 33342和碘化丙啶（PI）双染色显示，未分化的chES细胞发生细胞死亡，但外源性G6P可以挽救细胞。然而，通过PAS染色，我们发现分化的chES细胞减少了糖原储备。此外，分化的chES细胞表达了更高水平的GLUT1、HK1和PFK mRNA，而对照CEF细胞的GYS mRNA水平保持相似。这些数据表明，未分化的chES细胞继续以G6P为代价从葡萄糖合成糖原，而分化的chES细胞的糖原储备减少，这表明糖原的数量指示了chES细胞状态。
Cellular energy metabolism correlates with cell fate, but the metabolic properties of chicken embryonic stem (chES) cells are poorly understood. Using a previously established chES cell model and electron microscopy (EM), we found that undifferentiated chES cells stored glycogen. Additionally, undifferentiated chES cells expressed lower levels of glucose transporter 1 (GLUT1) and phosphofructokinase (PFK) mRNAs but higher levels of hexokinase 1 (HK1) and glycogen synthase (GYS) mRNAs compared with control primary chicken embryonic fibroblast (CEF) cells, suggesting that chES cells direct glucose flux towards the glycogenic pathway. Moreover, we demonstrated that undifferentiated chES cells block gluconeogenic outflow and impede the accumulation of glucose-6-phosphate (G6P) from this pathway, as evidenced by the barely detectable levels of pyruvate carboxylase (PCX) and mitochondrial phosphoenolpyruvate carboxykinase (PCK2) mRNAs. Additionally, cell death occurred in undifferentiated chES cells as shown by Hoechst 33342 and propidium iodide (PI) double staining, but it could be rescued by exogenous G6P. However, we found that differentiated chES cells decreased the glycogen reserve through the use of PAS staining. Moreover, differentiated chES cells expressed higher levels of GLUT1, HK1 and PFK mRNAs, while the level of GYS mRNA remained similar in control CEF cells. These data indicate that undifferentiated chES cells continue to synthesize glycogen from glucose at the expense of G6P, while differentiated chES cells have a decreased glycogen reserve, which suggests that the amount of glycogen is indicative of the chES cell state.