Experimental study on the synergistic action of retinol on leukemia inhibitory factor (LIF) maintaining undifferentiated state of murine embryonic stem cells / 解放军医学杂志

ABSTRACT

Objective To investigate the synergistic effect of retinol and leukemia inhibitory factor (LIF) on maintaining pluripotency of murine embryonic stem cells (mESCs). Methods The mESCs were randomly divided into two groups, and they were all cultured with the same LIF concentration (100U/ml). Retinol was added into mESCs medium in experimental group, and retinol was not added in control group. 14 days after culturing, the changes in cell morphology were observed under an inverted microscope with alkaline phosphatase staining. The mRNA expression of Oct4 and Nanog was analyzed by RT-PCR, and the proteins for Oct4 and Nanog were determined by Western-blot for the assessment of the pluripotency of mESCs. Results 14 days after culturing, 78% of mESCs in the experimental group formed good cell colonies with good stereoscopic appearance, and they showed strong alkaline phosphatase (AKP) staining. While in the control group, only 35% of mESCs were in undifferentiated mode, and the cells were weakly positive with AKP staining. The quantity of undifferentiated mESCs in experimental group (7.02?0.34) was higher than that in control group (4.28?0.37, P