Objective To explore an optional condition to induce mouse embryonic stem cell(ESC) to differentiate into endothelial cells so as to provide seedcells for tissue engineered vascular. Methods The embryos from one pregnant 12.5days mouse was harvested to culture the mouse embryonic fibroblasts(MEF). The ESC was reanimated by common method, and used to cultured into embryoid body(EB) in vitro. The EB which was used to induce into endothelial cells was divided into two groups. The EB was cultured in the EB medium with 3ng/ml transforming growth factor β1, 50 ng/ml vascular endothelial cell growth factor and 1 μmol/L potent and selective inhibitor of activin receptorlike kinase receptors in experimental group. The EB was cultured in the EB medium in the control group. After 14 days, RTPCR and immunohistochemistry were used to detect vWF and CD34, to analyze the morphology and type of the differentiated cells fromESC. Results The primary MEF had a high proliferation activity. At the 3rdday, the fusion rate of MEF was about 90% with a fusiform shape. The cells was fusiform shape and arranged compactly with fullness of nucleus and 2-3 entoblasts. The 3rd5th generations EB was polygonal with fullness of cytoplasm and 3-4 entoblasts. ESC could maintain undifferentiated state, and the cells unit lookedlike bird nest with smooth margin; the cells was small at size and b refractivity with high rate of nuclein and rapid proliferation. At 3 days of dropculture, EB can seen grossly and at 3 days of suspension, large and transparent EBformed. EB was spread radiately with an intensive adhesion at the 2nd day. In experimental group, many round cells was differentiated around EB from the 4thday to the 7th day, and form tubular structures from the 10th day to the 14th day. The vWF and CD34 were expressed. In control group, EB could not form tubularstructures, and the vWF and CD34 were not expressed. Conclusion ESC can differentiate into endothelial cells under some conditions, and form vessellike structure under condition culture, which can provide sources of seed cells for tissue engineered vessel.
Citation: XIONG Jixin,LIU Zhaoxuan,LIU Xiaochun,et al.. INDUCED DIFFERENTIATION OF MOUSE EMBRYONIC STEM CELL INTO ENDOTHELIALCELL IN VITRO. Chinese Journal of Reparative and Reconstructive Surgery, 2007, 21(9): 994-998. doi: Copy