Germ Cell Differentiated Into Hepatic Cells And The Study Of NF-κB And UBC9

Objective:To study the expression of NF-κB and UBC9during the differentiation of mouse PGCs to liver cells and the development of embryo liver, to explore the molecular mechanism of differentiation of stem cell into hepatocytes.Methods:The primordial germ cells(PCCs)from the gonadal ridges of the mouse embryos were cultured and proliferated in vitro.20μg/Lβ-nerve growth factor (NGF),0.5g/L newborn mice liver extract and fetal hepatocyte supernatant were added into mice embryonic stem cells respectively, the morphological changes of the mouse PGCs were observed; Expression of alpha-1anti-trypsin(AAT), NF-κB protein by immunofluores-cence, Expression of UBC9in the differentiation process of the mouse PGCs into hepatocyte were assayed by immunohistochemistry. And Real-Time PCR.assayed NF-κB and UBC9relative expression of mRNA. Expression of NF-κB and UBC9in the liver during embryonic development were detected by immunohistochemistry.Results:After the induction of20μg/Lβ-NGF,0.5g/L newborn mice liver extract and fetal liver cell supernatant, some PGCs became round or oval in12days which showed that PGCs begin to differentiate into hepatocyte. The round differentiated cells were in positive expression of alpha1anti-trypsin and the expression of NF-κB gradually increased in the process of differentiation, the positive NF-κB expression transferred from the cytoplasm into the nucleus with the liver cells mature. UBC9expression gradually increases. Real-Time PCR results showed that in the process of differentiation NF-κB expression increases, the expression of UBC9is increased. In fetal liver the expression of NF-κB in hepatic cells were also gradually increased, with the hepatocyte maturation the expression of NF-κB in nucleus were much more significant; UBC9expression in11.5d-14.5d gradually strengthen,18.5d, liver cell matures, UBC9almost no expression in the nucleus.Conclusion:20μg/Lβ-NGF,0.5g/L newborn mice liver extract and fetal liver cell supernatant could induce mice PGCs differentiation into liver cells. During the differentiation expression of NF-κB and UBC9gradually increased; And in the process of fetal liver development with liver cells to mature liver cells, the NF-κB expression gradually strengthened, UBC9expression enhanced first, then gradually weakened.