抗坏血酸对动态培养体系中mESCs扩增特性的影响

Ascorbic acid promoted ex vivo proliferation of mESCs in dynamic culture

在动态培养体系的培养过程中向培养基添加抗坏血酸后,可延迟小鼠胚胎干细胞(Mouse embryonic stem cells,mESCs)内与老化相关的β-半乳糖苷酶活性显著升高的时间,从培养6 d延迟至10 d;同时,实验组细胞内的活性氧水平均低于对照组,两组之间在培养8~9 d时呈显著性差异;在培养8 d时,实验组mESCs的总细胞扩增倍数和SSEA-1+细胞的扩增倍数分别为53.9±1.5和51.9±6.3,显著高于对照组的41.3±0.9和40.1±3.9。可见,抗坏血酸可显著降低细胞内活性氧水平,明显改善动态培养时mESCs的老化状态,促进mESCs的体外扩增。该结果为解决胚胎干细胞规模化培养过程中细胞老化问题提供了一种解决思路。

In this research,the growth characteristic of mouse embryonic stem cells(mESCs)in dynamic culture was studied.With the addition of ascorbic acid,the significantly increasing time ofβ-galactosidase activity which was related to senescence delayed from day 6 to day 10.At the same time,intracellular reactive oxygen species level in the experimental group was lower than that in the control group,and there was a significant difference between the two groups at day 8-day 9.Further analysis showed that the total expansion fold of mESCs and the expansion fold of SSEA-1+cells were 53.9±1.5 and 51.9±6.3 at day 8,respectively,both of which were significantly higher than those in control group without ascorbic acid,respectively(41.3±0.9 and 40.1±3.9).These data proved that ascorbic acid can reduce the level of intracellular reactive oxygen species significantly,improve the aging state of mESCs obviously and promote ex vivo expansion of mESCs.The research provides a new way to solve the aging problem of ESCs in the process of enlarged culture.