条件培养液改进小鼠胚胎干细胞嵌合体制备效率的观察

目的:探讨如何通过改善小鼠胚胎干细胞(embryonic stem cell,ES)的培养条件来维持其未分化状态,提高嵌合体的制备效率.方法:以普通ES细胞培养液为对照,以TX-WES培养液为ES细胞条件培养液分别对15代以上的未工程化ES细胞和糖皮质激素受体(GR)工程化ES细胞进行培养并筛选,通过囊胚注射制作嵌合体来比较2种不同培养液培养ES细胞后嵌合体制备效率的差别.结果:未工程化ES细胞以普通培养液培养其嵌合率为25.0%,以TX WES培养液培养其嵌合率提高到53.8%(P＜ 0.05);GR工程化ES细胞以普通培养液培养其嵌合率为15.0%,以TX-WES培养液培养其嵌合率提高到52.3%(P＜0.05).结论:应用TX-WES培养液改善小鼠ES细胞的培养条件可提高嵌合体的制备效率,该方法为基因剔除小鼠研究的顺利开展奠定了良好的基础.

Conditional medium improving obtainment of mouse embryonic stem cell chimeras

Objective:To increase the efficiency of mouse embryonic stem cell(ES) chimeras production through improving the culture condition to maintain undifferentiated state of ES cells.Methods: With traditional ES cells medium being control, TX-WES medium was used as conditional ES cells medium to culture non-targeting and GR targeting ES cell lines, which was passaged over 15 times. Chimeras was produced by blastocyst injection and the chimeras production efficiency between the 2 different media was compared. Results: Compared with traditional ES cells medium, Non-targeting ES cells cultured with TX-WES medium had a higher production of chimeras (53.8% versus 25.0%, P < 0.05); similarly, compared with traditional ES cells medium, GR targeting ES cells cultured with TX-WES medium also had a higher production of chimeras (52.3% versus 15.0%, P < 0.05). Conclusion: The results indicate that the chimeras production efficiency of mouse ES cells can be increased by improving ES cells' culture condition with TX-WES medium.