一种制备胎鼠成纤维细胞饲养层的简易方法

The simple method of reparation for MEF feeder layer cells

采用新的传代及MMC（Mytomycin-C,MMC）处理方法,以求简化繁琐的饲养层制备过程,对MEF（Mouse embryonic fibroblast,MEF）细胞采用胰酶消化（充分去除多余消化液）后直接接种的方法传代.MEF细胞经MMC处理后,用PBS充分洗去MMC继续培养2-3 h后做饲养层.结果表明：新的传代方法对MEF细胞的增殖及细胞形态无明显影响,新方法制备的饲养层具有促进ES细胞（Embryonic stem cell,ES）增殖以及抑制ES细胞分化的作用,而且得到了典型的ES细胞样集落,AKP呈阳性.研究结论表明,采用新的细胞传代方法及饲养层制备方法有效地简化了饲养层的制备过程,缩短了饲养层的准备时间.

In order to simplify the complicated process of preparation for feeder layers, the new method was used in subculture and MMC-treatment. The MEF cells were directly fed after being digested by trypsin/ EDTA solution. MMC-treated MEF cells were continuously cultured for 2 - 3 h after PBS-washed. The results showed that the new method of subculture did not affect the growth and morphology of the MEF cells; and the MEF feeder layers by using the new method promoted the ES cells proliferation and inhibited differentiation. The typical ES cell-like colonies were also obtained. AKP dyeing was strongly positive. Therefore, the new methods simplified the preparation process and shortened the preparation time of feeder layer cells.