6-磷酸葡萄糖对鸡类囊胚发育的影响

Effect of glucose-6-phosphate on development of blastocyte-like structure

由鸡Ⅹ期胚盘获得的类囊胚包含禽类胚胎干细胞,且具有较强的增殖与分化能力。体外培养12、24、36 h的类囊胚细胞S tathm in基因的表达高于Ⅹ期胚盘细胞,证实增殖的细胞为禽类胚胎干细胞。超微结构观察表明,类囊胚细胞在早期增殖过程中以卵黄蛋白为能量的主要来源,当卵黄蛋白消耗殆尽,则细胞出现大量死亡。胚胎干细胞培养液(ESM)能够促进类囊胚细胞的增殖却不能减少死亡。在ESM基础上添加5 mm o l/L 6-磷酸葡萄糖(G 6P),不仅各期细胞增殖显著,且24 h细胞死亡数的比例(D/T)由23%下降到5.6 7%(P<0.0 5),3 6 h细胞D/T由3 0%下降到1 5.6 3%(P<0.0 5)。结果表明,在ESM基础上添加5 mm o l/L G 6P可促进类囊胚细胞的增殖,并明显改善能量匮乏导致的细胞死亡。

The chicken blastocyte-like structure （BLS） derived from stage X embryo consists of blastodermal cells among which there are embryonic stem cells. These cells present high level of proliferation and can differentiate into various cell types from three embryo layers. Although Stathmin was expressed in both embryo and BLS, its expression level was significantly higher in the BLS, suggesting that the ES cells of BLS extremely proliferated after culturing. The percentage of the dead cells to the total number of cells （D/T） in the BLS was high after 24 h when cultured with basic medium （BM）, which may be explained by the sequential decreasing yolk protein stores which is the main endogenous energy source in the cytoplasm of the cells. In order to ameliorate this situation, embryonic stem cell medium （ESM） was used in culture. As a result, the cells of group ESM proliferated dramatically contrasting with those of group BS, but their death ratio after 24 h didn＇t decrease. Whereas, when glucose-6-phosphate （G6P） （5 mmol/L） was added in, the D/ T ratio of these cells decreased from 23% to 5.67% （P〈0.05） in 24 h and from 30% to 15.63% （P〈0.05） in 36 h. In conclusion, the medium of embryonic stem cell with G6P promotes cell proliferation and decreases cell death efficiently.