Role of Leukemia Inhibitory Factor in the Effect of Co-Culture on Preimplantation Embryo Develpement

Lee, Kyu Sup; Kim, Sang Woo; Na, Yong Jin; Lee, Young Ah; Kim, Ha Jung; Jang, Sung Kyu

Abstract

OBJECTIVE
To assess the effect of recombinant human leukemia inhibitory factor on in vitro development of 1-cell ICR mouse embryo. MATERIALS AND METHOD: ICR mice were superovulated with PMSG/hCG and 1-cell stage mouse embryos were recruited. 1-cell mouse embryo were cocultured on human oviductal cells in a CO2 incubator (coculture group) and were cultured on 0.4% BSA+HTF media (control group). And anti-hLIF Ab was added the cocultured group in a different concentration (1pg, 10pg, 100pg, 1ng) and developmental rate was compaired to the control group, and rhLIF was added to the preincubated 0.4% BSA+HTF media in a different concentration (2000U, 1000U, 100U, 10U) and its developmental rate was compaired to group which was cultured on 0.4% BSA+HTF media only. RESULT: 1. The cleavage rate of 2-cell mouse embryo co-cultured with human tubal epithelial cell was significantly higher than that of cultured with media alone (HTF with 0.4% BSA) (p<0.05). 2. When LIF antibody was added to the medium with human tubal epitherlial cell, the mouse embryo could not cleave more than 2-cell in 1 ng of LIF antibody, and less than 1 ng, the cleavage rate was lower than cultured without LIF antibody group(p<0.05). 3. Two cell blocked ICR mouse embryos were developed into four cells under LIF(p<0.05), but no further development was observed.
CONCLUSIONS
These results shows that LIF enhances the development of preimplantation embryo, and when rhLIF is applicated in vitro, it has positive effects on the development of early mouse embryo and can help overcoming the two-cell block.